Purification of oligosaccharides or mixtures of oligosaccharides

Abstract

The present invention relates to methods for purifying oligosaccharides or mixtures of oligosaccharides from fermentation broths, to the products of these methods and to the use of the products of these methods.

Description

Technical Field

[0001] The present invention relates to methods for purifying oligosaccharides or oligosaccharide mixtures from solutions, the products of these methods and the uses of the products of these methods. Background Art

[0002] So far, oligosaccharides are receiving more and more attention. Oligosaccharides are very diverse in chemical structure and are composed of various monosaccharides, such as glucose, galactose, N-acetylglucosamine, xylose, rhamnose, fucose, mannose, N-acetylneuraminic acid, N-acetylgalactosamine, galactosamine, glucosamine, glucuronic acid, galacturonic acid. Oligosaccharides are widely distributed in all living organisms and play an important role in multiple physiological and pathological processes, such as cell metastasis, signal transduction, intercellular adhesion, inflammation and immune response. The economic production of these oligosaccharides is extremely important in order to fully benefit from its biological advantages. The example of this type of oligosaccharide is the milk oligosaccharide (MO) in mammalian milk, referred to as mammalian milk oligosaccharide or MMO, and the milk oligosaccharide in human milk, referred to as human milk oligosaccharide (HMO) (Usashima T. et al., 2011). Many of these MOs contain fucose residues, galactose residues, N-acetylglucosamine or N-acetylneuraminic acid residues at their non-reducing ends. The importance of MO to animal and human infant nutrition is directly related to its biological activity, including protecting newborns from pathogens and supporting the development of infant immune systems and cognitive abilities. In addition, MO is used as a substrate for beneficial bacteria such as Bifidobacterium or Lactobacillus. It is further known that MO acts as a bait to reduce the risk of infection by bacterial and viral pathogens, which adhere to human cells by binding to the surface glycoproteins of these cells. In addition, various MOs have anti-inflammatory effects and act as immunomodulators (e.g., reducing the risk of food allergies). A variety of synthetic methods have been developed, ranging from extraction, chemical synthesis to enzymatic synthesis. These methods are currently least used, while biotechnology fermentative production is currently underway and commercialized. Methods for producing oligosaccharides are reviewed by Lu et al. (2021), Faijes et al. (2019), Kruschitz et al. (2020), Ghosh et al. (2020), Vera et al. (2021), Walsh et al. (2020), Li et al. (2020), Li and Ye (2020) and are well known to those skilled in the art. For all production methods, the final oligosaccharides need to be purified.

[0003] manual SUMMARY OF THE INVENTION

[0005] It is an object of the present invention to provide a process by which an oligosaccharide or oligosaccharide mixture can be purified from a solution, preferably in an efficient, time- and cost-effective manner, and producing the desired oligosaccharide or oligosaccharide mixture in high quality, high purity and good yield, wherein the desired oligosaccharide or oligosaccharide mixture obtained after purification has an ash content of ≤10% based on total dry solids and a lead content of less than 0.1 mg / kg dry solids, preferably less than 0.02 mg / kg dry solids, more preferably less than 0.01 mg / kg dry solids, an arsenic content of less than 0.2 mg / kg dry solids, preferably less than 0.05 mg / kg dry solids, more preferably less than 0.02 mg / kg dry solids, a cadmium content of less than 0.1 mg / kg dry solids, preferably less than 0.01 mg / kg dry solids and / or a mercury content of less than 0.5 mg / kg dry solids, preferably less than 0.1 mg / kg dry solids, more preferably less than 0.005 mg / kg dry solids.

[0006] According to the present invention, this and other objects are achieved by providing a method for purifying said oligosaccharides from a solution. The method comprises passing a solution containing the desired oligosaccharide through a solution containing H + The first cation exchange of the cation exchange resin in the form of a cation exchange resin is carried out, thereby converting the salt in the solution into acid completely or partially, and passing the solution containing the salt converted into acid completely or partially through a cation exchange resin containing Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + Preferably, the second cation exchange resin is Na + form. The solution containing oligosaccharides is any one of a biocatalytic reaction solution, a chemical synthesis solution, a cell culture, or a process stream of the above method, wherein the oligosaccharides are produced by the biocatalytic reaction solution, the chemical synthesis solution, or by cells cultured in the cell culture. Preferably, the cell culture is fermentation. The present invention also provides purified oligosaccharides prepared by the above method. In addition, the present invention provides a purified oligosaccharide mixture containing the purified oligosaccharides prepared by the above method. Upon reading the present invention, other benefits of the teachings of the present invention will be apparent to those skilled in the art.

[0007] definition

[0008] The words used in this specification to describe the present invention and its various embodiments should be understood not only to have their generally defined meanings, but also to include structures, materials, or actions that go beyond the generally defined meanings through special definitions in this specification. Therefore, if an element can be understood to include more than one meaning in the context of this specification, its use in the claims must be understood to be general to all possible meanings supported by the specification and the word itself.

[0009] The various aspects and embodiments of the present invention disclosed herein should not only be understood in the order and context specifically described in this specification, but also include any order and any combination thereof. Unless otherwise indicated, each embodiment described herein can be combined. All publications, patents and patent applications mentioned in this specification are incorporated herein by reference to the extent that each individual publication, patent or patent application is specifically and individually indicated to be incorporated by reference as a whole. Unless otherwise specifically indicated, all words used in the singular should be considered to include the plural, and vice versa. Unless otherwise defined, all technical and scientific terms used herein generally have the same meaning as those generally understood by those of ordinary skill in the art to which the present invention belongs. Typically, the nomenclature used herein and the laboratory procedures in cell culture, molecular genetics, organic chemistry and nucleic acid chemistry and hybridization described herein are those well known and commonly used in the art. Standard techniques are used for nucleic acid and peptide synthesis. Typically, enzymatic reactions and purification steps are carried out according to the manufacturer's instructions.

[0010] In this specification, embodiments of the present invention have been disclosed, and although specific terms have been employed, these terms are used in a descriptive sense only and not for the purpose of limitation, and the scope of the invention is set forth in the appended claims. It must be understood that the illustrated embodiments are set forth for illustrative purposes only and should not be construed as limiting the present invention. It will be apparent to those skilled in the art that changes, other embodiments, improvements, details, and uses consistent with the text and spirit disclosed herein may be made, and within the scope of this disclosure, the scope of this disclosure is limited only by the claims interpreted in accordance with patent law (including the doctrine of equivalents). In the claims that follow, the figure marks used to designate claim steps are provided for ease of description only and are not intended to imply any particular order for performing the steps, unless otherwise specifically stated.

[0011] Throughout this application, unless expressly stated otherwise, the features “synthesize,” “synthetic,” and “synthesize” are used interchangeably with the features “produce,” “produce,” and “produce,” respectively. Throughout this application, unless expressly stated otherwise, the expressions “can… <verb>” and “can… <verb>” are preferably replaced with the active voice of the verb, and vice versa. For example, the expression “can express” is preferably replaced with “express,” and vice versa, i.e., “express” is preferably replaced with “can express.” Throughout this document and its claims, the verbs “comprise,” “have,” and “contain,” and their conjugations, are used in their non-limiting sense to mean that the items following the word are included, but items not specifically mentioned are not excluded. The verb “consisting essentially of means that a solution or composition as defined herein may contain additional components in addition to the specifically identified components, which additional components do not alter the unique characteristics of the invention. The additional compounds may be unavoidable by-products, for example, by-products produced during the production of the oligosaccharides or oligosaccharide mixtures of the present invention, and compounds introduced into the process stream from which the oligosaccharides or oligosaccharide mixtures are recovered but cannot be removed therefrom. The term "essentially consisting of" with respect to spray-dried powders includes spray-dried powders that contain at least 80%, at least 85%, at least 90%, at least 93%, at least 95%, or at least 98% by weight of an oligosaccharide mixture relative to the dry matter of the spray-dried powder. The term "essentially consisting of" is also used for spray-dried powders, process streams, and solutions containing an oligosaccharide mixture. Throughout this document and its claims, unless otherwise specifically stated, the verbs "comprise," "have," and "contain," and their variations, may preferably be replaced by "consisting of" (and variations thereof) or "essentially consisting of" (and variations thereof), and vice versa. In addition, mentioning element by the indefinite article " a (a) " or " a kind of (an) " does not exclude the possibility that there is more than one element, unless the context clearly requires the presence of one and only one element. Therefore, the indefinite article " a " or " a kind of " usually means " at least one / a kind of ". Throughout the document and claims, unless otherwise clearly stated, the article " a " and " a kind of " are preferably replaced by " at least one ", more preferably " at least two ", even more preferably by " at least three ", even more preferably by " at least four ", even more preferably by " at least five ", even more preferably by " at least six ", most preferably by " at least two ". When used in combination with a numerical value (for example, " about 10 ") or a range (for example, " about x to about y "), the word " about " or " approximately " preferably means that the value or range is interpreted as being as accurate as the method for measuring it. If no error margin is specified, then when used in combination with a numerical value, the expression " about " or " approximately " is interpreted as having the same rounding as a given value.In this document and in the claims, unless otherwise indicated, the expression "x to y" (wherein x and y represent numerical values) refers to a numerical range where x is the lower limit of the range and y is the upper limit of the range. In this document, x and y are also included in the range.

[0012] The terms "sialic acid," "N-acetylneuraminate," "N-acylneuraminate," "N-acetylneuraminic acid," and "Neu(n)Ac molecule" are used interchangeably and refer to an acidic sugar with a nine-carbon backbone, including but not limited to Neu4Ac; Neu5Ac; Neu4,5Ac2; Neu5,7Ac2; Neu5,8Ac2; Neu5,9Ac2; Neu4,5,9Ac3; Neu5,7,9Ac3; Neu5,8,9Ac3; Neu4,5,7,9Ac4; Neu5,7,8,9Ac4; Neu4,5,7,8,9Ac5; Neu5Gc, and 2-keto-3-deoxymannosyl-octanone (KDO).

[0013] As used herein, the term "monosaccharide" refers to a sugar that cannot be broken down into simpler sugars by hydrolysis, is classified as an aldose, ketose, deoxysugar, deoxyaminosugar, uronic acid, aldonic acid, ketoalonic acid, aldaric acid, or sugar alcohol, and contains one or more hydroxyl groups per molecule. A monosaccharide is a sugar that contains only one simple sugar.

[0014] As used herein, the term "phosphorylated monosaccharide" refers to a phosphorylated monosaccharide. Examples of phosphorylated monosaccharides include, but are not limited to, glucose-1-phosphate, glucose-6-phosphate, glucose-1,6-bisphosphate, galactose-1-phosphate, fructose-6-phosphate, fructose-1,6-bisphosphate, fructose-1-phosphate, glucosamine-1-phosphate, glucosamine-6-phosphate, N-acetylglucosamine-1-phosphate, mannose-1-phosphate, mannose-6-phosphate, or fucose-1-phosphate.

[0015] The terms "activated monosaccharide", "nucleotide activated sugar", "nucleotide-sugar", "activated sugar", "nucleoside" or "nucleotide donor" are used interchangeably herein and refer to the activated form of a monosaccharide. Examples of activated monosaccharides include, but are not limited to, UDP-N-acetylglucosamine (UDP-GlcNAc), UDP-N-acetylgalactosamine (UDP-GalNAc), UDP-N-acetylmannosamine (UDP-ManNAc), UDP-glucose (UDP-Glc), UDP-galactose (UDP-Gal), GDP-mannose (GDP-Man), UDP-glucuronate, UDP-galacturonate, UDP-2-acetamido-2,6-dideoxy-L-arabinose-4-hexulose, UDP-2-acetamido-2,6-dideoxy-L-lyxose-4-hexulose, UDP-N-acetyl-L-rhamnosamine (UDP-L-RhaNAc or UDP-2-acetamido-2,6-dideoxy-L- Nucleotide sugars include UDP-N-acetylfucosamine (UDP-L-FucNAc or UDP-2-acetamido-2,6-dideoxy-L-galactose), UDP-N-acetyl-L-pneNAc or UDP-2-acetamido-2,6-dideoxy-L-talose), UDP-N-acetylmuramic acid, UDP-N-acetyl-L-quinolosamine (UDP-L-QuiNAc or UDP-2-acetamido-2,6-dideoxy-L-glucose), GDP-L-quinolosamine, CMP-sialic acid (CMP-Neu5Ac or CMP-N-acetylneuraminic acid), GDP-fucose (GDP-Fuc), GDP-rhamnose, UDP-xylose, and CMP-KDO. Nucleotide sugars serve as glycosyl donors in glycosylation reactions. Glycosylation reactions are catalyzed by glycosyltransferases.

[0016] As used herein, the term "glycosyltransferase" refers to an enzyme that is capable of catalyzing the transfer of a sugar moiety from a donor to a specific acceptor, thereby forming a glycosidic bond. The donor may be a precursor as defined herein. Glycosyltransferases have been described as being classified into different sequence-based families using nucleotide diphosphate sugars, nucleotide monophosphate sugars, and sugar phosphates, as well as related proteins (Campbell et al., Biochem. J. 326, 929-939 (1997)) and are available on the CAZy (Carbohydrate Active Enzymes) website (www.cazy.org). As used herein, the glycosyltransferase may be selected from a list including, but not limited to, fucosyltransferase, sialyltransferase, galactosyltransferase, glucosyltransferase, mannosyltransferase, N-acetylglucosaminyltransferase, N-acetylgalactosaminyltransferase, N-acetylmannosaminyltransferase, xylosyltransferase, glucuronyltransferase, galacturonosyltransferase, glucosaminyltransferase, N-glycolylneuraminic acid transferase, rhamnosyltransferase, N-acetylrhamnosyltransferase, UDP-4-amino-4,6-dideoxy-N-acetyl-β-L-altrosamine transaminase, UDP-N-acetylglucosamine enolpyruvyltransferase, and fucosamine transferase.

[0017] As used herein, the term "disaccharide" refers to a sugar polymer containing two simple sugars (i.e., monosaccharides). Examples of disaccharides include lactose (Gal-β1,4-Glc), lactulose (Gal-β1,4-Fru), allolactose (Gal-β1,6-Glc), lactose-N-disaccharide (Gal-β1,3-GlcNAc), N-acetyllactosamine (Gal-β1,4-GlcNAc), LacDiNAc (GalNAc-β1,4-GlcNAc), N-acetylgalactosamine (Gal ... The 247 amino acids were 1,4-nitro-1,2-dimethylaminopropanediol (1,4-d-glucose), 2,6-nitro-1,2-dimethylaminopropanediol (1,4-d-glucose), 2,7 ...

[0018] As used herein and as generally understood in the prior art, the term "oligosaccharide" refers to a sugar polymer containing a small amount, typically three to twenty, preferably three to ten simple sugars (i.e., monosaccharides). The oligosaccharides used in the present invention can be linear structures or can include branches. The bond between two sugar units (e.g., glycosidic bond, galactosidic bond, glucosidic bond, etc.) can be expressed as, for example, 1,4, 1->4, or (1-4), which are used interchangeably herein. For example, the terms "Gal-b1,4-Glc", "Gal-β1,4-Glc", "b-Gal-(1->4)-Glc", "β-Gal-(1->4)-Glc", "Galβ1-4-Glc", "Gal-b(1-4)-Glc" and "Gal-β(1-4)-Glc" have the same meaning, i.e., the β-glycosidic bond connects the carbon-1 of galactose (Gal) to the carbon-4 of glucose (Glc). Each monosaccharide can be in a cyclic form (e.g., a pyranose or furanose form). The bonds between the individual monosaccharide units can include α1->2, α1->3, α1->4, α1->6, α2->1, α2->3, α2->4, α2->6, β1->2, β1->3, β1->4, β1->6, β2->1, β2->3, β2->4, and β2->6. Oligosaccharides can contain both α-glycosidic bonds and β-glycosidic bonds, or can contain only α-glycosidic bonds or only β-glycosidic bonds. The term "polysaccharide" refers to a compound composed of a large number (usually more than twenty) of glycosidically linked monosaccharides.Examples of oligosaccharides include, but are not limited to, Lewis-type antigenic oligosaccharides, mammalian (including human) milk oligosaccharides, O-antigens, Enterobacterial common antigens (ECA), glycan chains present in lipopolysaccharides (LPS), oligosaccharide repeating units present in capsular polysaccharides, peptidoglycans (PG), amino sugars, antigens of the human ABO blood group system, animal oligosaccharides (which are preferably selected from the group consisting of N-glycans and O-glycans), plant oligosaccharides (which are preferably selected from the group consisting of N-glycans and O-glycans), ), sialylated oligosaccharides, neutral (uncharged) oligosaccharides, negatively charged oligosaccharides, fucosylated oligosaccharides, oligosaccharides containing N-acetylglucosamine, oligosaccharides containing lactose-N-disaccharide, oligosaccharides containing N-acetyllactosamine, sialylated oligosaccharides containing N-acetylglucosamine, sialylated oligosaccharides containing N-acetylglucosamine, neutral (uncharged) oligosaccharides containing N-acetylglucosamine, negatively charged oligosaccharides containing N-acetylglucosamine, oligosaccharides containing N-acetylglucosamine, Fucosylated oligosaccharides containing acetylglucosamine, non-fucosylated oligosaccharides containing N-acetylglucosamine, sialylated oligosaccharides containing lactose-N-biose, neutral (uncharged) oligosaccharides containing lactose-N-biose, negatively charged oligosaccharides containing lactose-N-biose, fucosylated oligosaccharides containing lactose-N-biose, non-fucosylated oligosaccharides containing lactose-N-biose, sialylated oligosaccharides containing N-acetyllactosamine, neutral (uncharged) oligosaccharides containing N-acetyllactosamine (Uncharged) oligosaccharides, negatively charged oligosaccharides containing N-acetyllactosamine, fucosylated oligosaccharides containing N-acetyllactosamine, non-fucosylated oligosaccharides containing N-acetyllactosamine, chitosan, oligosaccharides comprising chitosan, heparin precursor (heparosan), chondroitin sulfate, glycosaminoglycan oligosaccharides, heparin, heparan sulfate, dermatan sulfate, hyaluronan, hyaluronic acid, keratan sulfate, glucopyranosylsucrose (erlose, Glc-α1,4-Glc-α1,2-Fru); lactulose-N-triose II (GlcNAc-β1,3-Gal-β1,4-Fru); lactulose-N-tetraose; lactulose-N-neotetraose; uridine diphosphate galactose (globotriose).

[0019] The terms "negatively charged oligosaccharide" or "acidic oligosaccharide" are used interchangeably and refer to oligosaccharides having a negative charge. In a preferred embodiment, the negatively charged oligosaccharide is a sialylated oligosaccharide. As used herein, "sialylated oligosaccharide" is understood to mean an oligosaccharide containing a negatively charged sialic acid, i.e., an oligosaccharide having one or more sialic acid residues. It has an acidic nature. Some examples are 3'SL (3'-sialyllactose, Neu5Ac-α2,3-Gal-β1,4-Glc), 3'-sialyllactosamine, 6'SL (6'-sialyllactose, Neu5Ac-α2,6-Gal-β1,4-Glc), 8'SL (8'-sialyllactose, Neu5Ac-α2,8-Gal-β1,4-Glc), 3,6-disialyllactose (Neu5Ac-α2,3-(Neu5Ac-α2,6)-Gal-β1,4-Glc), 6,6'- Disialyl lactose (Neu5Ac-α2,6-Gal-β1,4-(Neu5Ac-α2,6)-Glc), 8,3-disialyl lactose (Neu5Ac-α2,8-Neu5Ac-α2,3-Gal-β1,4-Glc), 6'-sialyllactosamine, 3'-sialyllactosamine, oligosaccharides containing 6'-sialyllactose, SGG hexose (Neu5Acα-2,3Galβ-1,3GalNacβ-1,3Galα-1,4Galβ-1,4Gal), sialylated tetrasaccharides Sugar, sialylated pentasaccharide, sialylated lactose-N-triosaccharide, sialylated lactose-N-tetraosaccharide, sialylated lactose-N-neotetraosaccharide, LSTc (Neu5Ac-α2,6-Gal-β1,4-GlcNAc-β1,3-Gal-β1,4-Glc), LSTd (Neu5Ac-α2,3-Gal-β1,4-GlcNAc-β1,3-Gal-β1,4-Glc), monosialyllactose-N-hexaosaccharide, disialyllactose-N-hexaosaccharide I, monosialyllactose-N-neohexaose I, Monosialyllactose-N-neohexaose II, disialyllactose-N-neohexaose, disialyllactose-N-tetraose, disialyllactose-N-hexaose II, sialyllactose-N-tetraose a (LSTa, Neu5Ac-α2,3-Gal-β1,3-GlcNAc-β1,3-Gal-β1,4-Glc), disialyllactose-N-hexaose I, sialyllactose-N-tetraose b (LSTb, Gal-β1,3-(Neu5Ac-α2,6)-GlcNAc-β1,3-Gal-β1,4-Glc)4-Glc), 3'-sialyl-3-fucosyllactose, fucodisialyllactose-N-hexaose, disialylmonofucosyllactose-N-neohexaose, monofucosylmonosialyllactose-N-octaose (sialyl Lea), sialyllactose-N-fucohexaose II, disialyllactose-N-fucopentaose II, monofucosyldisialyllactose-N-tetraose, Neu5Ac-a2,3-Gal-b1,4-GlcNAc-b1,3-Gal, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-KDO-lactose, 3'-KDO-lactosamine, 3'-KDO-6'sialyllactose, 3'KDO-8'sialyllactose Lactose, KDO-2,3Galβ-1,3GalNacβ-1,3Galα-1,4Galβ-1,4Gal, KDO-2,3Galβ-1,3GlcNacβ-1,3Galβ-1,4Glc, KDO-2,3Galβ-1,4GlcNacβ-1,3Galβ-1,4Glc, 3'-KDO-3-fucosyllactose, Neu5Ac-a2,8-Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-sialyl-2'-fucosyllactose, 6'-sialyl-2'-fucosyllactose, 6'-sialyl-3-fucosyllactose, 3'-sialyl Neu5Ac-a2,6-(Neu5Ac-a2,3-)Gal-b1,4-Glc, 3'-sialyl-3-fucosyllactosamine, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc, 6'-sialyl lactose-N-disaccharide, 3'-sialyl lactose-N-disaccharide, Neu5Ac-a2,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,3-Gal- b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-( Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-) Glc, Neu5Ac-a2,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a 1,2-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b 1,3-Gal-b1,4-Glc, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5A c-a2,6-(Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-( Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3 -GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc and oligosaccharides with one or several sialic acid residues, including but not limited to: oligosaccharide moieties of gangliosides selected from GM3 (3' sialyllactose, Neu5Acα-2,3Galβ-4Glc) and oligosaccharides containing the GM3 motif, GD3 Neu5Acα-2,8Neu5Acα-2,3Galβ-1,4Glc GT3(Neu5Acα-2,8Neu5Acα-2,8Neu5Acα-2,3Galβ-1,4Glc); GM2GalNAcβ-1,4(Neu5Acα-2,3)Galβ-1,4Glc, GM1 Galβ-1,3GalNAcβ-1,4(Neu5Acα-2,3)Galβ-1,4Glc, GD1a Neu5Acα-2,3Galβ-1,3GalNAcβ-1,4(Neu5Acα-2,3)Galβ-1,4Glc, GT1aNeu5Acα-2,8Neu5Acα-2,3Galβ-1,3GalNAcβ-1,4(Neu5Acα-2,3)Galβ-1,4Glc, GD2 GalNAcβ-1,4(Neu5Acα-2,8Neu5Acα2,3)Galβ-1,4Glc, GT2 GalNAcβ-1,4(Neu5Acα-2,8Neu5Acα-2,8Neu5Acα2,3)Galβ-1,4Glc,GD1b,Galβ-1,3GalNAcβ-1,4(Neu5Acα-2,8Neu5Acα2,3)Galβ-1,4Glc,GT1b Neu5Acα-2,3Galβ-1,3GalNAcβ-1,4(Neu5Acα-2,8Neu5Acα2,3)Galβ-1,4Glc, GQ1bNeu5Acα-2,8Neu5Acα-2,3Galβ-1,3GalNAc β-1,4(Neu5Acα-2,8Neu5Acα2,3)Galβ-1,4Glc, GT1c Galβ-1,3GalNAcβ-1,4(Neu5Acα-2,8Neu5Acα-2,8Neu5Acα2,3)Galβ-1,4Glc, GQ1c Neu5Acα-2,3Galβ-1,3GalNAcβ-1,4(Neu5Acα-2,8Neu5Acα-2,8Neu5Acα2,3)Galβ-1,4Glc,GP1c Neu5Acα-2,8Neu5Acα-2,3Galβ-1,3GalNAcβ-1,4(Neu5Acα-2,8Neu5Acα-2,8Neu5Acα2,3)Galβ-1,4Glc, GD1aNeu5Acα-2,3Galβ-1,3(Neu5Acα-2,6)GalNAcβ-1,4Galβ-1,4Glc, Fucosyl-GM1 Fucα-1,2Galβ-1,3GalNAcβ-1,4(Neu5Acα-2,3)Galβ-1,4Glc; all of these can be extended to produce the corresponding gangliosides by reacting the above oligosaccharide moieties with ceramide or synthesizing the above oligosaccharides on ceramide.

[0020] "Charged oligosaccharides" are oligosaccharide structures containing one or more negatively charged monosaccharide subunits, including N-acetylneuraminic acid (Neu5Ac), commonly known as sialic acid, N-glycolylneuraminic acid (Neu5Gc), glucuronic acid esters, galacturonic acid esters, and 2-keto-3-deoxymannose-octanone acid (KDO). Charged oligosaccharides are also referred to as acidic oligosaccharides. Sialic acid belongs to the family of derivatives of neuraminic acid (5-amino-3,5-dideoxy-D-glycero-D-galactose-nonyl-2-keto acid). Neu5Gc is a derivative of sialic acid, which is formed by the hydroxylation of the N-acetyl group at C5 of Neu5Ac. In contrast, neutral (uncharged) oligosaccharides are non-sialylated oligosaccharides and therefore do not contain acidic monosaccharide subunits. Neutral oligosaccharides include uncharged fucosylated oligosaccharides containing one or more fucose subunits in their glycan structure and uncharged non-fucosylated oligosaccharides lacking any fucose subunits. Other examples of charged oligosaccharides are sulfated chitosan and deacetylated chitosan.

[0021] As used herein, the terms "neutral oligosaccharide" and "uncharged" oligosaccharide are used interchangeably and, as generally understood in the art, refer to oligosaccharides that do not have a negative charge derived from a carboxylic acid group. Examples of such neutral oligosaccharides are 2'-fucosyllactose (2'FL), 3-fucosyllactose (3FL), 4-fucosyllactose (4FL), 6-fucosyllactose (6FL), 2',3-difucosyllactose (diFL), lacto-N-triose II (LN3, GlcNAcβ1-3Galβ1-4Glc), lacto-N-tetraose (LNT, Galβ1-3GlcNAcβ1-3Galβ1-4Glc), lacto-N-neotetraose (LNnT, Galβ1-4GlcNAcβ1-3Galβ1-4Glc), lacto-N-fucopentaose I, lacto-N-neotetraose (LNnT, Galβ1-4GlcNAcβ1-3Galβ1-4Glc), lacto-N-fucopentaose I, lacto-N-neotetraose (LNnT, Galβ1-4GlcNAcβ1-3Galβ1-4Glc), lacto-N-fucopentaose II ... Algae pentasaccharide I, lactose-N-fucopentasaccharide II, lactose-N-fucopentasaccharide III, lactose-N-fucopentasaccharide V, lactose-N-fucopentasaccharide VI, lactose-N-neofucopentasaccharide V, lactose-N-difucosylhexaose I, lactose-N-difucosylhexaose II, Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc- a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1 ,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,4-(Fuc-a1,2-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, monofucosyllactose N -hexasaccharide-III, difucosyllactose N-hexaose (a), 6'-galactosyl lactose, 3'-galactosyl lactose, lactose-N-hexaose, lactose-N-neohexaose, p-lactose-N-hexaose, p-lactose-N-neohexaose, difucosyl-lactose-N-hexaose, difucosyl-lactose-N-neohexaose, trifucosyllactose N-hexaose, a1,3-galactosyl-3-fucosyllactose, Gal-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc3-)Glc, 2-fucosyllactulose, 3-fucosyl-N-acetyllactosamine, 2'-fucosyl-N-acetyllactosamine, difucosyl-N-acetyllactosamine, 4-fucosyllactose N-disaccharide, 2'-fucosyllactose N-disaccharide, difucosyllactose N-disaccharide and GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, GlcNAc-b1 ,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, lactose-N-pentaose (LN5), lactose-N-neopentaose, p-lactose-N-pentaose, p-lactose-N-neopentaose, lactose-N-novel pentasaccharide I, lactose-N-heptasaccharide (LN7), lactose-N-neoheptasaccharide, p-lactose-N-neoheptasaccharide, p-lactose-N-heptasaccharide, lactose-N-octaose (LNO), lactose-N-novoheptasaccharide, isolactose Lactose-N-octaose, p-lactose-N-octaose, allolactose-N-neooctaose, novel lactose-N-neooctaose, p-lactose-N-neooctaose (pLNnO), allolactose-N-nonaose, novel lactose-N-nonaose, lactose-N-nonaose (LN9), lactose-N-decaose, allolactose-N-decaose, novel lactose-N-decaose, lactose-N-neodecaose, p-lactose-N-neodecaose (pLNnD), a1,3-galactosyl Lactose-N-neotetraose, GlcNAc-b1,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, GlcNAc-b1,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc and GlcNAc-b1,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc.

[0022] As used herein and as generally understood in the art, a 'fucosylated oligosaccharide' is an oligosaccharide that carries a fucose residue. Such a fucosylated oligosaccharide is a sugar structure comprising at least three monosaccharide subunits linked to each other by glycosidic bonds, wherein at least one of the monosaccharide subunits is fucose. A fucosylated oligosaccharide may contain more than one fucose residue, for example, two, three, or more. A fucosylated oligosaccharide may be a neutral oligosaccharide or a charged oligosaccharide, for example, it may also contain a sialic acid structure. Fucose may be linked to other monosaccharide subunits comprising glucose, galactose, or GlcNAc via α-glycosidic bonds, including α-1,2, α-1,3, α-1,4, or α-1,6 bonds.Examples include 2'-fucosyllactose (2'FL), 3-fucosyllactose (3FL), 4-fucosyllactose (4FL), 6-fucosyllactose (6FL), difucosyllactose (diFL), lactose-N-fucopentaose I (LNFP I), Gal-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (Gal-LNFP I), GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (GalNAc-LNFP I), lactose-N-fucopentaose II (LNFP II), lactose-N-fucopentaose III (LNFP III), lactose-N-fucopentaose V (LNFP V), lactose-N-fucopentaose VI (LNFP VI), lactose-N-neofucopentaose I, lactose-N-difucohexaose I (LDFH I), lactose-N-difucohexaose II (LDFH II), monofucosyllactose-N-hexaose III (MFLNH III), difucosyllactose-N-hexaose (a), difucosyllactose-N-hexaose, difucosyllactose-N-neohexaose, trifucosyllactose-N-hexaose, a1,3-galactosyl-3-fucosyllactose, Gal-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, 2-fucosyllactulose, 3-fucosyl-N-acetyllactosamine, 2'-fucosyl-N-acetyllactosamine, difucosyl-N-acetyllactosamine, 4-fucosyllactose Glycosyllactose-N-disaccharide, 2'-fucosyllactose-N-disaccharide, difucosyllactose-N-disaccharide and GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, 3'-sialyl-2'-fucosyllactose, 6'-sialyl-2'-fucosyllactose, 6'-sialyl-3-fucosyllactose, 3'-sialyl-3-fucosyllactose, disialylmonofucosyllactose-N-neohexaose, monofucosylmonosialyllactose-N-octaose (sialic acid Lea), sialyllactose-N-fucohexaose II, disialyllactose-N-fucopentaose II, monofucosyldisialyllactose-N-tetraose.

[0023] Mammalian milk oligosaccharides include oligosaccharides present in milk found at any stage of lactation, including colostrum from humans and colostrum from mammals including, but not limited to, cows (Bos Taurus), sheep (Ovisaries), goats (Capra aegagrus hircus), Bactrian camels (Camelus bactrianus), horses (Equus feruscaballus), pigs (Sus scropha), dogs (Canis lupus familiaris), Hokkaido brown bears (Ursus arctosyesoensis), polar bears (Ursus maritimus), Japanese black bears (Ursus thibetanus japonicus), striped skunks (Mephitis mephitis), hooded seals (Cystophora cristata), Asian elephants (Elephas maximus), African elephants (Loxodonta africana), giant anteaters (Myrmecophaga tridactyla), bottlenose dolphins (Tursiops truncates), northern minke whales (Balaenoptera acutorostrata), Eugene wallaby (Macropuseugenii), red kangaroo (Macropus rufus), common brushtail possum (Trichosurus vulpecula), koala (Phascolarctos cinereus), eastern quoll (Dasyurus viverrinus), platypus (Ornithorhynchus anatinus).As used herein, "mammalian milk oligosaccharide" or "MMO" refers to oligosaccharides such as, but not limited to, 3-fucosyllactose, 2'-fucosyllactose, 6-fucosyllactose, 2',3-difucosyllactose, 2',2-difucosyllactose, 3,4-difucosyllactose, 6'-sialyllactose, 3'-sialyllactose, 3,6-disialyllactose, 6,6 '-Disialyl lactose, 8,3-disialyl lactose, 3,6-disialyl lactose-N-tetraose, lactose-N-tetraose, lactose-N-neotetraose, lactose-N-fucopentaose II, lactose-N-fucopentaose I, lactose-N-fucopentaose III, lactose-N-fucopentaose V, lactose-N-fucopentaose VI, sialyllactose-N-tetraose c, sialyllactose-N -tetraose b, sialyllactose-N-tetraose a, lactose-N-difucohexaose I, lactose-N-difucohexaose II, lactose-N-hexaose, lactose-N-neohexaose, para-lactose-N-hexaose, monofucosylmonosialyllactose-N-tetraose c, monofucosylpara-lactose-N-hexaose, monofucosyllactose-N-hexaose III, isomeric fucosylated lactose-N-hexaose III, isomeric fucosylated lactose-N-hexaose I, sialyllactose-N-hexaose, sialyllactose-N-neohexaose II, difucosyl lactose-N-hexaose, difucosyllactose-N-hexaose, difucosyllactose-N-hexaose a, difucosyllactose-N-hexaose c, galactosylated chitosan, fucosylated oligosaccharides, neutral oligosaccharides and / or sialylated oligosaccharides.

[0024] The term "human milk oligosaccharide" or "HMO" refers to oligosaccharides found in human breast milk (including premature human milk, colostrum and full-term human milk). HMOs include fucosylated oligosaccharides, non-fucosylated neutral oligosaccharides and sialylated oligosaccharides (see, e.g., Chen X., Chapter 4: Human Milk Oligosaccharides (HMOs): Structure, Function, and Enzyme-Catalyzed Synthesis in Adv. Carbohydr. Chem. Biochem. 72, 113 (2015). Examples of HMOs include 3-fucosyllactose, 2'-fucosyllactose, 2',3-difucosyllactose, 6'-sialyllactose, 3'-sialyllactose, LN3, lacto-N-tetraose, lacto-N-neotetraose, lacto-N-fucopentaose II, lacto-N-fucopentaose I, lacto-N-fucopentaose III, lacto-N-fucopentaose V, lacto-N-fucopentaose VI, sialic acid Lactose-N-tetraose c, sialyllactose-N-tetraose b, sialyllactose-N-tetraose a, difucosyllactose-N-tetraose, lactose-N-hexaose, lactose-N-difucohexaose I, lactose-N-difucohexaose II, disialyllactose-N-tetraose, fucosyllactose-N-hexaose, difucosyllactose-N-hexaose, fucodisialyllactose-N-hexaose, disialyllactose-N-hexaose.

[0025] "Recombinant" means a genetically engineered DNA prepared by transplanting or splicing a gene from one species into a cell of a host organism of a different species. This DNA becomes a part of the host's genetic makeup and is replicated. As used herein, the terms "recombinant" or "transgenic" or "metabolic engineered" or "genetically engineered" are used interchangeably relative to cells or host cells and indicate that cells replicate heterologous nucleic acids, or express peptides or proteins encoded by heterologous nucleic acids (i.e., sequences that are "exogenous to the cell" or sequences that are "exogenous to the position or environment in the cell"). Such cells are described as being transformed with at least one heterologous or exogenous gene, or as being transformed by introducing at least one heterologous or exogenous gene. Recombinant or metabolically engineered cells can contain genes not found in the natural (non-recombinant) form of the cell. Recombinant cells can also contain genes found in the natural form of the cell, wherein the genes are modified and reintroduced into the cell by artificial means. The term also includes cells containing endogenous nucleic acids of the cell that have been modified or whose expression or activity has been modified without removing the nucleic acid from the cell; such modifications include those obtained by gene replacement, promoter replacement, site-specific mutation, and related techniques. Thus, a "recombinant polypeptide" is a polypeptide produced by a recombinant cell. The term also encompasses cells that have been modified by removing endogenous nucleic acids of the cell by common techniques well known to those skilled in the art (e.g., knocking out a gene).

[0026] Protein or polypeptide sequence information and functional information can be provided by a comprehensive resource of protein sequence and annotation data, such as the Universal Protein Resource (UniProt) (www.uniprot.org) (Nucleic Acids Res. 2021, 49 (D1), D480-D489). UniProt includes a specialized and richly organized protein database known as the UniProt Knowledge Base (UniProtKB), as well as the UniProt Reference Cluster (UniRef) and the UniProt Archive (UniParc). The UniProt identifier (UniProt ID) is unique for each protein present in the database. Throughout the application, the sequence of a polypeptide is represented by a UniProt ID. Unless otherwise indicated, the UniProt ID of the protein corresponds to its sequence version 01 present in the UniProt database (www.uniprot.org) version release 2021_03 and accessed on June 9, 2021. It will be understood by those skilled in the art that for the databases used herein (including UniProt), the content of each database is fixed and does not change at each release. When the content of a particular database changes, the particular database receives a new published version with a new publication date. All published versions of each database and their corresponding publication dates and the specific content annotated at these specific publication dates are available and known to those skilled in the art.

[0027] As used herein, the term "cell productivity index (CPI)" refers to the mass of oligosaccharide produced by cells divided by the mass of cells produced in culture.

[0028] Term " mammary cell " as used herein generally refers to mammalian mammary epithelial cell, mammalian mammary epithelial luminal cell or mammalian epithelial alveolar cell or its any combination.Term " mammary sample cell " as used herein generally refers to the mammalian cell with the phenotype / genotype similar to natural mammalian mammary cell (or substantially similar) but derived from mammalian non-mammary cell source.Such mammalian mammary sample cell can be engineered to remove at least one unwanted genetic component and / or include at least one mammalian mammary cell typical predetermined genetic construct.The limiting examples of mammalian mammary sample cell can include mammalian mammary epithelial cell, mammalian mammary epithelial luminal cell, the mammalian non-mammary cell of one or more characteristics of the cell showing mammalian mammary cell lineage, or its any combination.Other limiting examples of mammalian mammary sample cell can include the phenotype similar to natural mammalian mammary cell (or substantially similar), or more particularly the mammalian cell with the phenotype similar to natural mammalian mammary epithelial cell (or substantially similar). Mammalian cells having a phenotype similar to (or substantially similar to) natural mammalian mammary cells or mammalian mammary epithelial cells or showing at least one feature similar to (or substantially similar to) natural mammalian mammary cells or mammalian mammary epithelial cells can include natural mammalian cells (e.g., derived from mammary cell lineages or non-mammary cell lineages) that express at least one milk component. As used herein, the term "non-mammary cells" can generally include any mammalian cells of non-mammary lineages. In the context of the present invention, non-mammary cells can be any mammalian cells that can be engineered to express at least one milk component. Non-limiting examples of such non-mammary cells include hepatocytes, blood cells, kidney cells, umbilical cord blood cells, epithelial cells, epidermal cells, muscle cells, fibroblasts, mesenchymal cells, or any combination thereof. In some cases, molecular biology and genome editing techniques can be engineered to eliminate, silence, or weaken numerous genes simultaneously.

[0029] The terms "cultivation," "cell culture," and "incubation" are used interchangeably; these terms include the medium in which the cells are cultured or fermented or incubated, the medium components, the cells themselves, and the oligosaccharides produced by the cells in the whole fermentation broth, i.e., inside the cells (intracellular) as well as outside the cells (extracellular).

[0030] As used herein, term " biomass " refers to suspension, precipitation or insoluble material derived from fermentation cells, such as intact cells, broken cells, cell fragments, cell walls, phospholipids, cell membranes, proteins, protein fragments, polysaccharides, polynucleotides and other large organic compounds produced by cells. Biomass can be in suspension and / or in solution. As used herein, biomass is also understood to be non-complex biomass, which generally has low or no cell organization that is converted into special or complex structure. Non-limiting examples are single cells, cell pairs, cell agglomerates, oligocellular or multicellular structures, cell layers, biofilms. Non-complex biomass can have three-dimensional structure, such as cambium or biofilm or cells attached to reactor or incubator (incubator) surface. As used herein, biomass is also understood to be complex biomass, wherein the complex biomass has complex structure in nature, typically complex three-dimensional structure and comprising hundreds, thousands, tens of thousands, but more typically hundreds of thousands or millions or more cell structures, typically with various cell types of different specificities. Non-limiting examples are higher plants or animals with a body visible to the naked eye, organs and tissues including bones and flesh or plant parts such as fruit, vegetables, straw, bagasse, hay, wood, timber. Complex biomass can be a source of non-complex biomass, for example cell lines are typically derived from tissues or organs but do not maintain a complex structure in culture.

[0031] As used herein, the term "cell dry weight" or "CDW" refers to the dry weight of biomass in grams per liter of sample after removal of water. The procedure for measuring CDW is disclosed in the working examples below.

[0032] The terms "biocatalytic reaction solution" and "enzymatic synthesis reaction" are used interchangeably and refer to a mixture in which oligosaccharides are produced enzymatically. The mixture may comprise one or more enzymes, one or more precursors, and one or more receptors as defined herein, present in a buffer solution and incubated at a temperature and for a period of time such that oligosaccharides can be produced by the one or more enzymes using the one or more precursors and the one or more receptors in the mixture. The mixture may also comprise i) cells that produce one or more enzymes, one or more precursors, and / or one or more receptors as defined herein and used in the biocatalytic reaction to produce oligosaccharides, and ii) a buffer solution or culture or incubation medium in which the cells are cultured or incubated.

[0033] The term "chemical synthesis solution" is understood to mean a mixture in which oligosaccharides are chemically produced. The chemical synthesis solution may comprise one or more reactants, one or more intermediate compounds, and one or more by-products, which are incubated at a certain temperature and for a certain period of time to enable the production of oligosaccharides in the solution through one or more chemical reactions. The chemical synthesis solution may also comprise one or more catalysts that accelerate or slow down the synthesis reactions in the chemical synthesis solution.

[0034] The term "any process stream" is understood to mean any solution present, used, or generated in any step of the purification process of the present invention. Examples of such process streams include, but are not limited to, inlet solution, outlet solution, influent, effluent, eluent, eluate, fluid, waste solution, buffer, solvent, alcohol, acid, base, lysate, filtrate, extract.

[0035] The terms "reactor" and "incubator" refer to a container filled with a culture, incubation, chemical synthesis solution, or biocatalytic reaction solution. Examples of reactors and incubators include, but are not limited to, microfluidic devices, well plates, tubes, shake flasks, fermentors, bioreactors, processing vessels, cell culture incubators, and CO2 incubators. The reactors and incubators can each vary in size from laboratory-scale to large-scale industrial sizes.

[0036] The term "purified" refers to a material that is substantially or essentially free of components that interfere with the activity of biomolecules. For cells, sugars, nucleic acids and polypeptides, the term "purified" refers to a material that is substantially or essentially free of the following components, which are usually accompanied by the material found in its native state. Typically, the purity of the purified sugars, oligosaccharides, proteins or nucleic acids of the present invention is at least about 50%, 55%, 60%, 65%, 70%, 75%, 80% or 85%, usually at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99.0%, which is measured by band intensity on a silver stained gel or other methods for determining purity. Purity or homogeneity can be indicated by many methods well known in the art, such as polyacrylamide gel electrophoresis of a protein or nucleic acid sample, which is then visualized when stained. For some purposes, high resolution will be required and HPLC or similar purification methods will be used. For disaccharides and oligosaccharides, purity can be determined using methods such as, but not limited to, thin layer chromatography, gas chromatography, NMR, HPLC, capillary electrophoresis, or mass spectrometry. In addition, the terms "contaminants" and "impurities" herein preferably refer to particles, cells, cell components, metabolites, cell debris, proteins, peptides, amino acids, nucleic acids, glycolipids, and / or endotoxins that may be present in an aqueous medium (e.g., a culture, incubation, chemical synthesis solution, or a biocatalytic reaction solution).

[0037] As used herein, the term "clarification" refers to the act of treating an aqueous medium (e.g., a culture, incubation, chemical synthesis solution, or biocatalytic reaction solution) to remove suspended particles and contaminants (e.g., cells, cell components, insoluble metabolites, and debris) from the production process that may interfere with the final purification of the oligosaccharide solution, oligosaccharide, or oligosaccharide mixture. Such treatment may be performed in a conventional manner by centrifugation, flocculation, flocculation with optional sonication, gravity filtration, microfiltration, foam separation, or vacuum filtration (e.g., by a Celite filtration system that may include Celite filtration). TM Ceramic filter with filter aid).

[0038] As used herein, the term "protein-free oligosaccharide solution" refers to an oligosaccharide solution from a culture, incubation, chemical synthesis solution or a biocatalytic reaction solution that has been treated to remove substantially all proteins and any associated impurities, such as amino acids, peptides, endotoxins, glycolipids, RNA and DNA, from the process, which may interfere with the final purification of the oligosaccharide solution from the process. Protein, preferably substantially all protein, can be removed by ion exchange chromatography, affinity chromatography, ultrafiltration and size exclusion chromatography in a conventional manner. Preferably, the protein-free oligosaccharide solution is a clarified oligosaccharide solution. According to the present invention, the term "purifying oligosaccharide solution from culture" means gathering, collecting or reclaiming an oligosaccharide solution from the culture medium of a cell and / or its growth.

[0039] "Purified oligosaccharide solution" comprises an oligosaccharide or a mixture of oligosaccharides dissolved in an aqueous medium. The aqueous medium is a solvent comprising water. In some embodiments, the aqueous medium is pure water. In other embodiments, the medium comprises water and trace amounts of one or more organic solvents. In some such embodiments, the medium comprises less than 1 wt % (weight percent) organic solvent. In some embodiments, the medium comprises less than 0.1 wt % organic solvent. In some embodiments, the medium comprises less than 0.01 wt % organic solvent. In some embodiments, the medium comprises less than 0.001 wt % organic solvent. In some embodiments, the medium comprises less than 0.0001 wt % organic solvent.

[0040] In some embodiments, the oligosaccharide solution comprises trace amounts of one or more organic solvents. In some such embodiments, the purified oligosaccharide solution comprises less than 1% by weight organic solvent. In some embodiments, the purified oligosaccharide solution comprises less than 0.1% by weight organic solvent. In some embodiments, the purified oligosaccharide solution comprises less than 0.01% by weight organic solvent. In some embodiments, the purified oligosaccharide solution comprises less than 0.001% by weight organic solvent. In some embodiments, the purified oligosaccharide solution comprises less than 0.0001% by weight organic solvent.

[0041] In some embodiments, the purity of the oligosaccharides in the solution is measured based on total dry solids. In this article, total dry solids includes particles, cells, cell components, metabolites, cell debris, proteins, peptides, amino acids, nucleic acids, glycolipids, lipids, endotoxins, salts, vitamins, heavy metals and carbohydrates.

[0042] As used herein, the term "precursor" refers to a substance that is taken up or synthesized by a cell to specifically produce an oligosaccharide according to the present invention. In this sense, a precursor can be an acceptor as defined herein, but can also be another substance, a metabolite, which is first modified in the cell as part of the biochemical synthesis pathway of an oligosaccharide. As used herein, the term "precursor" should also be understood as a compound that participates in a chemical synthesis reaction, incubation or biocatalytic reaction (i.e., an enzymatic reaction) as part of an oligosaccharide metabolic pathway to produce another compound (e.g., an intermediate or acceptor as defined herein). As used herein, the term "precursor" should also be understood as a donor for use by glycosyltransferases as part of an oligosaccharide metabolic pathway to modify an acceptor as defined herein with a sugar moiety in a glycosidic bond. Examples of such precursors include receptors as defined herein, and / or dihydroxyacetone, glucosamine, N-acetylglucosamine, N-acetylmannosamine, galactosamine, N-acetylgalactosamine, galactosyl lactose, phosphorylated sugars or sugar phosphates such as, but not limited to, glucose-1-phosphate, galactose-1-phosphate, glucose-6-phosphate, fructose-6-phosphate, fructose-1,6-diphosphate, mannose-6-phosphate, mannose-1-phosphate, glycerol-3-phosphate, glyceraldehyde-3-phosphate, dihydroxyacetone. Acetone-phosphate, glucosamine-6-phosphate, N-acetylglucosamine-6-phosphate, N-acetylmannosamine-6-phosphate, N-acetylglucosamine-1-phosphate, N-acetylneuraminic acid-9-phosphate and nucleotide activated sugars, such as nucleotide diphosphate sugars and nucleotide monophosphate sugars as defined herein, such as, for example, UDP-glucose, UDP-galactose, UDP-N-acetylglucosamine, CMP-sialic acid, GDP-mannose, GDP-4-dehydro-6-deoxy-α-D-mannose, GDP-fucose.

[0043] Optionally, the cells used to produce oligosaccharides are transformed to contain and express at least one nucleic acid sequence encoding a protein selected from the group consisting of a lactose transporter, an N-acetylneuraminic acid transporter, a fucose transporter, a glucose transporter, a galactose transporter, and a nucleotide-activated sugar transporter, wherein the transporter internalizes precursors added to the culture medium for the synthesis of the oligosaccharides of the present invention.

[0044] As used herein, the term "acceptor" refers to a monosaccharide, disaccharide, or oligosaccharide, which can be modified by a glycosyltransferase. Examples of such acceptors include glucose, galactose, fructose, glycerol, sialic acid, fucose, mannose, maltose, sucrose, lactose, lacto-N-triose, lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), lacto-N-pentaose (LNP), lacto-N-neopentaose, p-lactose-N-pentaose, p-lactose-N-neopentaose, lacto-N-neopentaose, lacto-N-neopentaose, lacto-N-neopentaose, lacto-N-neopentaose, lacto-N-hexaose (LNH), lacto-N-neohexaose (LNnH), p-lactose-N-neohexaose (pLNnH), p-lactose-N-hexaose (pLNH), lacto-N-heptaose, lacto-N-neoheptaose, p-lactose-N-neoheptaose, p-lactose-N-heptaose, lacto-N-octaose (LNO), lacto-N-neooctaose, allolactose-N- Octasaccharide, p-lactose-N-octaose, allolactose-N-neooctaose, novel lactose-N-neooctaose, p-lactose-N-neooctaose, allolactose-N-nonaose, novel lactose-N-nonaose, lactose-N-nonaose, lactose-N-decaose, allolactose-N-decaose, novel lactose-N-decaose, lactose-N-neodecaose and oligosaccharides containing one or more N-acetyllactosamine units and / or one or more lactose-N-disaccharide units or intermediates converted into oligosaccharides, fucosylated and sialylated variants thereof, ceramide, N-acylated sphingosine, glucosylceramide, lactosylceramide, sphingosine, phytosphingosine, sphingosine synthon, peptide backbone with β-GlcNAc-Asn residues, glycoproteins with GlcNAc and Gal terminal residues, immunoglobulins.

[0045] As used herein, "Brix value" represents the sugar content of an aqueous solution. The Brix value can be expressed as a percentage (Brix percentage) or "Brix degree". Strictly speaking, the Brix value is the weight percentage of sucrose in a pure aqueous solution, and is therefore not applicable to solutions containing other solutes and / or solvents. However, the Brix value is easy to measure and is therefore commonly used in the art as an approximation of the total sugar content of sugar solutions other than pure sucrose solutions. When a purified oligosaccharide solution contains two or more different oligosaccharides, the "Brix value" as used herein represents the combined sugar content of the aqueous solution. The technology for measuring the Brix value is well known in the art. The dissolution of sugar in an aqueous solution changes the refractive index of the solution. Therefore, a properly calibrated refractometer can be used to measure the Brix value of a solution. Alternatively, the density of the solution can be measured and converted into a Brix value. A digital density meter can automatically perform this measurement and conversion, or a hydrometer or a pycnometer can be used.

[0046] As used herein, the terms "dry solids" and "dry matter" are used interchangeably and are further described in Example 1.

[0047] As used herein, the term "flow rate" refers to how quickly a solution (e.g., a solution comprising oligosaccharides of the present invention) passes through a resin in, for example, ion exchange, cation exchange, anion exchange, mixed bed ion exchange. Flow rate is expressed in BV / h. The terms "bed volume / hour," "bed volume / h," "BV / hour," and "BV / h" are used interchangeably. The terms "bed volume" or "BV" are used interchangeably and refer to the volume (m2) of the resin used in ion exchange, ion exchange chromatography, mixed bed ion exchange, cation exchange, anion exchange, 3 As used herein, the term "bed volume" is also understood to mean the minimum volume of solvent required to wet a defined amount of adsorbent within a column. This may vary depending on the nature of the adsorbent.

[0048] Ash content is a measure of the total amount of minerals and heavy metals present in a food or ingredient such as oligosaccharides, where mineral content is the specific inorganic component present in the food, such as Ca 2+ 、Na + , K + Mg 2+ , phosphate, sulfate and Cl -The ash content of a food is a measure of the amount of inorganic components present in the food, such as lead, arsenic, mercury, cadmium, zinc, manganese, copper, and iron. Ash is the inorganic residue remaining after water and organic matter are removed by heating in the presence of an oxidizing agent, and it provides a measure of the total amount of minerals and heavy metals in the food. Analytical techniques used to provide information about the total mineral and heavy metal content are based on the fact that analytes such as minerals and heavy metals can be distinguished from all other components (matrix) within the food or ingredient in some measurable way. The most widely used methods are based on the fact that minerals and heavy metals are not destroyed by heating and have low volatility compared to other food components. Three main types of analytical procedures for determining the ash content of foods are based on this principle: dry ashing, wet ashing, and low-temperature plasma dry ashing. The method selected for a particular analysis depends on the reason for the analysis, the type of food or ingredient being analyzed, and the available equipment. Ashing can also be used as a first step in preparing samples for analysis of specific minerals and heavy metals by atomic spectroscopy or various traditional methods described below. For sample preparation, a sample whose composition is representative of the component is selected to ensure that its composition has not changed significantly prior to analysis. For example, dried oligosaccharide samples are often hygroscopic, and the selected sample should be kept under dry conditions to avoid water absorption. Typically, a 1-10g sample is used for ash content analysis. Solid components are finely ground and then carefully mixed to facilitate the selection of a representative sample. Samples with high moisture content or those in solution are typically dried before ash analysis to prevent splashing during the ashing process. Other potential problems include contamination of the sample by minerals in the grinder, glassware, or crucible that come into contact with the sample during analysis. For the same reason, deionized water is used when preparing samples and for blank samples. The dry ashing procedure utilizes a high-temperature muffle furnace capable of maintaining temperatures between 500 and 600°C. Water and other volatile substances are evaporated, and organic matter is combusted in the presence of oxygen from the air to produce CO2, H2O, and N2. Most minerals are converted to oxides, sulfates, phosphates, chlorides, or silicates. Although most minerals and heavy metals have fairly low volatility at these high temperatures, some are volatile and may be partially lost, such as iron, lead, and mercury, for which ICP-MS analysis of the product is more suitable for quantification. Detailed Description of the Invention

[0050] According to a first aspect, the present invention provides a method for purifying oligosaccharides from a solution, wherein the solution comprising the oligosaccharides is a solution selected from the group consisting of a biocatalytic reaction solution, a chemical synthesis solution, a cell culture, and any process stream of a method as described herein, and wherein the oligosaccharides are produced by the biocatalytic reaction solution, the chemical synthesis solution, or by cells cultured in the cell culture. More specifically, the present invention provides a method for purifying oligosaccharides from a solution, wherein the solution comprising the oligosaccharides is a solution selected from the group consisting of a biocatalytic reaction solution, a chemical synthesis solution, a cell culture, and a fermentation as described herein, and wherein the oligosaccharides are produced by the biocatalytic reaction solution, the chemical synthesis solution, or by cells cultured in the cell culture or the fermentation. The method comprises passing the solution comprising the oligosaccharides through a stream comprising H + The method comprises the steps of: first cation exchange with a cation exchange resin in the form of a cation exchange resin, thereby converting the salt in the solution completely or partially into an acid, characterized in that the solution containing the salt completely or partially converted into an acid is passed through a cation exchange resin containing Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + In a preferred embodiment, the cation exchange resin in the second cation exchange is Na + In this paper, from the H + The eluate obtained from the first cation exchange of a cation exchange resin in the form of + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 +Alternatively, the method further comprises any one or more processes selected from the group consisting of homogenization, clarification, removal, concentration, centrifugation, decantation, dilution, pH adjustment in the range of 2 to 5, temperature adjustment, filtration, ultrafiltration, microfiltration, diafiltration, reverse osmosis, nanofiltration, dialysis, use of activated carbon or carbon, use of a solvent in the range of pH 2 to 5, use of an alcohol, use of an aqueous alcohol mixture, use of charcoal, tangential flow high performance filtration, tangential flow ultrafiltration, affinity chromatography, hydrophobic interaction chromatography, gel filtration, ligand exchange chromatography, column chromatography, cation exchange adsorption resin, use of an adsorbent material, vacuum evaporation, wiped film evaporation, falling film evaporation, pasteurization, enzymatic treatment, decolorization, drying, and dissolution in a liquid in the range of pH 2 to 5, and wherein any one or more of the processes is performed in the presence of H + The first cation exchange is carried out after the cation exchange resin in the form of Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + The second cation exchange resin is in the form of a cation exchange resin.

[0051] The present invention relates to a method for purifying oligosaccharides, said oligosaccharides being provided in a solution comprising said oligosaccharides.

[0052] In a preferred embodiment, the oligosaccharide is selected from the list comprising, consisting of, or consisting essentially of: fucosylated oligosaccharides, neutral (uncharged) oligosaccharides, negatively charged oligosaccharides, sialylated oligosaccharides, Lewis-type antigenic oligosaccharides, oligosaccharides containing N-acetylglucosamine, neutral (uncharged) oligosaccharides containing N-acetylglucosamine, oligosaccharides containing N-acetyllactosamine, oligosaccharides containing lactose-N-diose, oligosaccharides containing galactose, non-fucosylated neutral (uncharged) oligosaccharides, chitosan, oligosaccharides containing chitosan, heparin precursors, chondroitin sulfate, glycosaminoglycan oligosaccharides, oligosaccharides containing ... Sugar, heparin, heparan sulfate, dermatan sulfate, hyaluronan or hyaluronic acid, keratan sulfate, milk oligosaccharide, O-antigen, Enterobacterial common antigen (ECA), oligosaccharide repeating units present in capsular polysaccharides, peptidoglycan, amino sugars, antigens of the human ABO blood group system, animal oligosaccharides, plant oligosaccharides, glucopyranosylsucrose (Glc-a1,4-Glc-a1,2-Fru), lactulose-N-trisaccharide II (GlcNAc-b1,3-Gal-b1,4-Fru), lactulose-N-tetraose, lactulose-N-neotetraose, uridine diphosphate galactose.

[0053] In a more preferred embodiment, the oligosaccharide is selected from the group consisting of fucosylated oligosaccharides: 2'-fucosyllactose (2'FL), 3-fucosyllactose (3FL), 4-fucosyllactose (4FL), 6-fucosyllactose (6FL), 2',3-difucosyllactose (diFL), lactose-N-fucopentaose I (LNFP I), Gal-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (GalNAc-LNFP I), GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (GalNAc-LNFP I), lactose-N-neofucopentaose I (LNFP I), I), lactose-N-fucopentaose II (LNFP II), lactose-N-fucopentaose III (LNFP III), lactose-N-fucopentaose V (LNFP V), lactose-N-fucopentaose VI (LNFP VI), lactose-N-neofucopentaose V, lactose-N-difucohexaose I (LNDFH-I), lactose-N-difucohexaose II (LNDFH-II), Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-GlcNAc-b1,3-Ga l-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-G lc, Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1 ,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc、Fuc-a1,4-(Fuc-a1,2-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc、monofucosyllactose-N-hexaose-III、difucosyllactose-N-hexaose(a ), difucosyl-lactose-N-hexaose, difucosyl-lactose-N-neohexaose, trifucosyllactose-N-hexaose, a1,3-galactosyl-3-fucosyllactose, Gal-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc3-)Glc, 2-fucosyllactulose, 3-fucosyl-N-acetyllactosamine, 2'-fucosyl-N-acetyllactosamine, difucosyl-N-acetyllactosamine, 4-fucosyllactose-N-disaccharide, 2'-fucosyllactose-N-disaccharide, difucosyllactose-N-disaccharide, and GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc. In another more preferred embodiment, the oligosaccharide is a sialylated oligosaccharide selected from the group consisting of 3' sialyllactose (3'SL), 6' sialyllactose (6'SL), 8' sialyllactose (8'SL), 3,6-disialyllactose (Neu5Ac-α2,3-(Neu5Ac-α2,6)-Gal-β1,4-Glc), 6,6'-disialyllactose (Neu5Ac-α2,6-Gal-β1,4-(Neu5Ac-α2,6)-Glc), 8,3-disialyllactose (Neu5Ac-α2,8-Neu5Ac-α2 ,3-Gal-β1,4-Glc), 6'-sialyllactosamine, 3'-sialyllactosamine, sialyllactose-N-tetraose a (LSTa), sialyllactose-N-tetraose b (LSTb), sialyllactose-N-tetraose c (LSTc), sialyllactose-N-tetraose d (LSTd), disialyllactose-N-tetraose, disialyllactose-N-neotetraose, monosialyllactose-N-hexaose, disialyllactose-N-hexaose I, disialyllactose-N-hexaose II, monosialyllactose-N-neohexaose I, monosialyllactose-N-neohexaose II, disialyllactose Sialyl lactose-N-neohexaose, 3'-sialyl-3-fucosyllactose, fucodisialyl lactose-N-hexaose, disialyl monofucosyllactose-N-neohexaose, sialyl lactose-N-fucohexaose II, disialyl lactose-N-fucopentaose II, monofucosyl disialyl lactose-N-tetraose, Neu5Ac-a2,3-Gal-b1,4-GlcNAc-b1,3-Gal, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-KDO-lactose, 3'-KDO-lactosamine, 3'- KDO-6' sialyllactose, 3'KDO-8-sialyllactose, KDO-2,3Galβ-1,3GalNacβ-1,3Galα-1,4Galβ-1,4Gal, KDO-2,3Galβ-1,3GlcNacβ-1,3Galβ-1,4Glc, KDO-2,3Galβ-1,4GlcNacβ-1,3Galβ-1,4Glc, 3'-KDO-3-fucosyllactose, Neu5Ac-a2,8-Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-sialyl-2'-fucosyllactose, 6'-sialyl-2'-fucosyllactose, 6'-sialyl-3-fucosyllactose, 3'-sialyl-3-fucosyllactose, Neu5Ac-a2,6-(Neu5Ac-a2,3-)Gal-b1,4-Glc, 3'-sialyl-3-fucosyllactosamine, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc, 6'-sialyl lactose-N-disaccharide, 3'-sialyl lactose-N-disaccharide, Neu5Ac-a2,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6 -(Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,3-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac- a2,6-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcN Ac-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Ga l-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-)Ga l-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc and any combination thereof. In another more preferred embodiment, the oligosaccharide is a neutral (uncharged) oligosaccharide containing N-acetylglucosamine selected from the following list: lactose-N-triose II (LN3), lactose-N-tetraose (LNT), lactose-N-neotetraose (LNnT), 6'-galactosyl lactose, 4'-galactosyl lactose, 3'-galactosyl lactose, GlcNAc-b1,6-Gal-b1,4-Glc, lactose-N-hexaose (LNH), lactose-N- Neohexaose (LNnH), p-lactose-N-hexaose (pLNH), p-lactose-N-neohexaose (pLNnH), GlcNAc-b1,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, lactose-N-pentaose (LN5), lactose-N-neopentaose, p-lactose-N-pentaose, p-lactose-N-neopentaose, lactose-N-novopentaose I, lactose-N-heptasaccharide (LN7), lactose-N-neoheptasaccharide, p-lactose-N-neoheptasaccharide Heptaose, p-lactose-N-heptasaccharide, lactose-N-octaose (LNO), lactose-N-neooctaose, allolactose-N-octaose, p-lactose-N-octaose, allolactose-N-neooctaose, novel lactose-N-neooctaose, p-lactose-N-neooctaose (pLNnO), allolactose-N-nonasaccharide, novel lactose-N-nonasaccharide, lactose-N-nonasaccharide (LN9), lactose-N-decaose, allolactose-N-decaose, novel lactose-N-decaose, lactose-N-neodecasaccharide, p-lactose- N-neodecasaccharide (pLNnD), a1,3-galactosyl lactose-N-neotetraose, GlcNAc-b1,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, GlcNAc-b1,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc and GlcNAc-b1,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc. In another more preferred embodiment, the oligosaccharide is a milk oligosaccharide. In an even more preferred embodiment, the oligosaccharide is a mammalian milk oligosaccharide (MMO) as described herein. In another even more preferred embodiment, the oligosaccharide is a human milk oligosaccharide (HMO) as described herein. In another more preferred embodiment, the oligosaccharide is an animal oligosaccharide selected from N-glycans and O-glycans. In another more preferred embodiment, the oligosaccharide is a plant oligosaccharide selected from N-glycans and O-glycans. In the context of the present invention, N-glycans and O-glycans refer to oligosaccharide structures known to those skilled in the art, wherein the structures are not linked to proteins or peptides.

[0054] In another more preferred embodiment, the oligosaccharide is a sialylated oligosaccharide having at least one sialic acid group selected from the group consisting of Neu4Ac; Neu5Ac; Neu4,5Ac2; Neu5,7Ac2; Neu5,8Ac2; Neu5,9Ac2; Neu4,5,9Ac3; Neu5,7,9Ac3; Neu5,8,9Ac3; Neu4,5,7,9Ac4; Neu5,7,8,9Ac4; Neu4,5,7,8,9Ac5; Neu5Gc and 2-keto-3-deoxymannose-octanone (KDO). In another more preferred embodiment, the oligosaccharide is a sialylated oligosaccharide having one Neu5Ac (neuraminic acid) group. In another more preferred embodiment, the oligosaccharide is a sialylated oligosaccharide having two sialic acid groups. In another more preferred embodiment, the oligosaccharide is a sialylated oligosaccharide having two identical sialic acid groups. In an even more preferred embodiment, the oligosaccharide is a sialylated oligosaccharide having two Neu5Ac groups. In another more preferred embodiment, the oligosaccharide is a sialylated oligosaccharide having three or more sialic acid groups.

[0055] In another more preferred embodiment, the oligosaccharide is a sialylated milk oligosaccharide. In an even more preferred embodiment, the oligosaccharide is a sialylated mammalian milk oligosaccharide (MMO). In a most preferred embodiment, the oligosaccharide is a sialylated human milk oligosaccharide (HMO) selected from the group consisting of: 3' sialyllactose, 6' sialyllactose, sialyllactose-N-tetraose a, sialyllactose-N-tetraose b, sialyllactose-N-tetraose c, sialyllactose-N-tetraose d, disialyllactose-N-tetraose, disialyllactose-N-neotetraose, monosialyllactose-N-hexaose, disialyllactose-N-hexaose i, disialyllactose-N-neotetraose The present invention relates to a novel oligosaccharide comprising the following: lactose-N-hexaose II, monosialyllactose-N-neohexaose I, monosialyllactose-N-neohexaose II, disialyllactose-N-neohexaose, 3'-sialyl-3-fucosyllactose, fucodisialyllactose-N-hexaose, dilactose sialylomonofucosyl-N-neohexaose, sialyllactose-N-fucohexaose II, disialyllactose-N-fucopentaose II and monofucosyldisialyllactose-N-tetraose, and combinations thereof.

[0056] In another more preferred embodiment, the oligosaccharide is a fucosylated oligosaccharide having at least one fucose group. In another more preferred embodiment, the oligosaccharide is a fucosylated oligosaccharide having two fucose groups. In another more preferred embodiment, the oligosaccharide is a sialylated oligosaccharide having three or more fucose groups.

[0057] In another more preferred embodiment, the oligosaccharide is a fucosylated milk oligosaccharide. In an even more preferred embodiment, the oligosaccharide is a fucosylated mammalian milk oligosaccharide (MMO). In a most preferred embodiment, the oligosaccharide is a fucosylated human milk oligosaccharide (HMO) selected from the group consisting of 2'-fucosyllactose (2'FL), 3-fucosyllactose (3FL), 4-fucosyllactose (4FL), 6-fucosyllactose (6FL), 2', 3-difucosyllactose (diFL), lactose-N-fucopentaose I (LNFP I), lactose-N-neofucopentaose I (LNFP I), lactose-N-fucopentaose II (LNFP II), lactose-N-fucopentaose III (LNFP III), lactose-N-fucopentaose V (LNFP V) and lactose-N-fucopentaose V (LNFP V). Lactose-N-fucopentaose (LNFPVI), lactose-N-neofucopentaose (LNFPVI), lactose-N-difucohexaose (LNDFH-I), lactose-N-difucohexaose (LNDFH-II), difucosyl-lactose-N-hexaose, and difucosyl-lactose-N-neohexaose.

[0058] In another more preferred embodiment, the oligosaccharide is selected from the list comprising: the list consisting of or the list consisting essentially of: 2'-fucosyllactose (2'FL), 3-fucosyllactose (3FL), 4-fucosyllactose (4FL), 6-fucosyllactose (6FL), 2',3-difucosyllactose (diFL), lactose-N-fucopentaose I (LNFPI), Gal-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (GalNAc-LNFP I), lactose-N-neofucopentaose I (LNFP II), I), lactose-N-fucopentaose II (LNFP II), lactose-N-fucopentaose III (LNFP III), lactose-N-fucopentaose V (LNFP V), lactose-N-fucopentaose VI (LNFP VI), lactose-N-neofucopentaose V, lactose-N-difucohexaose I (LNDFH-I), lactose-N-difucohexaose II (LNDFH-II), Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-GlcNAc-b1,3-Ga l-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-G lc, Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b 1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc、Fuc-a1,4-(Fuc-a1,2-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc、monofucyl lactose-N-hexaose-III、difucosyllactose-N-hexaose(a ), difucosyl-lactose-N-hexaose, difucosyl-lactose-N-neohexaose, trifucosyllactose-N-hexaose, a1,3-galactosyl-3-fucosyllactose, Gal-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc3-)Glc, 2-fucosyllactulose, 3-fucosyl-N-acetyllactosamine, 2'-fucosyl-N-acetyllactosamine, difucosyl-N-acetyllactosamine, 4-fucosyllactose-N-disaccharide, 2'-fucosyllactose-N-disaccharide, difucosyllactose-N-disaccharide, GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, 3'-sialyllactose (3'SL), 6'-sialyllactose (6'SL), 8'sialyllactose (8'SL), 3,6-disialyllactose (Neu5Ac-α2,3-(Neu5Ac-α2,6)-Gal-β1,4-Glc ), 6,6'-disialyl lactose (Neu5Ac-α2,6-Gal-β1,4-(Neu5Ac-α2,6)-Glc), 8,3-disialyl lactose (Neu5Ac-α2,8-Neu5Ac-α2,3-Gal-β1,4-Glc), 6'-sialyllactosamine, 3'-sialyllactosamine, sialyllactose-N-tetraose a (LSTa), sialyllactose-N-tetraose b (LSTb), sialyllactose-N-tetraose c (LSTc), sialyllactose-N-tetraose d (LSTd), disialyl lactose-N-tetraose, disialyl lactose-N-neotetraose, monosialyllactose-N-hexaose, disialyl lactose Acid lactose-N-hexaose I, disialyl lactose-N-hexaose II, monosialyllactose-N-neohexaose I, monosialyllactose-N-neohexaose II, disialyl lactose-N-neohexaose, 3'-sialyl-3-fucosyllactose, fucodisialyl lactose-N-hexaose, disialylmonofucosyllactose-N-neohexaose, sialyl lactose-N-fucohexaose II, disialyl lactose-N-fucopentaose II, monofucosyldisialyl lactose-N-tetraose, Neu5Ac-a2,3-Gal-b1,4-GlcNAc-b1,3-Gal, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-G al, 3'-KDO-lactose, 3'-KDO-lactosamine, 3'-KDO-6'-sialyllactose, 3'KDO-8-sialyllactose, KDO-2,3Galβ-1,3GalNacβ-1,3Galα-1,4Galβ-1,4Gal, KDO-2,3Galβ-1,3GlcNacβ-1,3Galβ-1,4Glc, KDO-2,3Galβ-1,4GlcNacβ-1,3Galβ-1,4Glc, 3'-KDO-3-fucosyllactose, Neu5Ac-a2,8-Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-sialyl-2'-fucosyllactose, 6'-sialyl-2'-fucosyllactose, 6'-sialyl-3-fucosyllactose, 3'-sialyl-3-fucosyllactose, Neu5Ac-a2,6-(Neu5Ac-a2,3-)Gal-b1,4-Glc, 3'-sialyl-3-fucosyllactosamine, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc, 6'-sialyl lactose-N-disaccharide, 3'-sialyl-N-disaccharide, Neu5Ac-a2,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6- (Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,3-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a 2,6-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNA c-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal -b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-)Gal -b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-Glc,N eu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, lactose-N-triose II (LN3), lactose-N-tetraose (LNT), lactose-N-neotetraose (LNnT), 6'-galactosyl lactose, 4'-galactosyl lactose, 3'-galactosyl lactose, GlcNAc-b1,6-Gal-b1,4-Glc, lactose-N-hexaose (LNH), lactose-N-neohexaose (LNnH), p-lactose-N-hexaose (pLNH), p-lactose-N-neohexaose (pLNnH), GlcNAc-b1,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, lactose-N-pentaose (LN5), lactose-N-neopentaose, p-lactose-N-pentaose, p-lactose-N-neopentaose, lactose-N-neopentaose, lactose-I, lactose- N-heptasaccharide (LN7), lactose-N-neoheptasaccharide, p-lactose-N-neoheptasaccharide, p-lactose-N-heptasaccharide, lactose-N-octaose (LNO), lactose-N-neooctaose, allolactose-N-octaose, p-lactose-N-octaose, allolactose-N-neooctaose, novel lactose-N-neooctaose, p-lactose-N-neooctaose (pLNnO), allolactose-N-nonasaccharide, novel lactose-N-nonasaccharide, lactose-N-nonasaccharide (LN9), lactose-N-decaose, allolactose-N-decaose, novel lactose-N-decaose, lactose-N -neodecaose, p-lactose-N-neodecaose (pLNnD), a1,3-galactosyllactose-N-neotetraose, GlcNAc-b1,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, GlcNAc-b1,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc and GlcNAc-b1,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc.

[0059] In another preferred embodiment, the oligosaccharide is an oligosaccharide having a degree of polymerization (DP) of at least 3. In another preferred embodiment, the oligosaccharide is an oligosaccharide having a DP selected from the group consisting of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20. In another preferred embodiment, the oligosaccharide is an oligosaccharide having a DP selected from the group consisting of 3, 4, 5, 6, 7, 8, 9, and 10. In another preferred embodiment, the oligosaccharide is a trisaccharide. In another preferred embodiment, the oligosaccharide is a tetrasaccharide. In another preferred embodiment, the oligosaccharide is a pentasaccharide. In another preferred embodiment, the oligosaccharide is a hexasaccharide. In another preferred embodiment, the oligosaccharide is a heptasaccharide.

[0060] In another preferred embodiment, the oligosaccharide is a linear oligosaccharide. In an alternative preferred embodiment, the oligosaccharide is a branched oligosaccharide. In the context of the present invention, the oligosaccharide is preferably in free form, i.e. the oligosaccharide does not contain any protecting groups.

[0061] In another preferred embodiment, the oligosaccharide is an oligosaccharide comprising one or more sialic acid groups and one or more monosaccharide building blocks selected from the group consisting of fucose, galactose, glucose, xylose, mannose, N-acetylglucosamine, N-acetylgalactosamine, rhamnose, glucuronic acid, galacturonic acid, and N-acetylmannosamine. In a more preferred embodiment, the oligosaccharide is a sialylated oligosaccharide that is also fucosylated. In another more preferred embodiment, the oligosaccharide is a non-fucosylated sialylated oligosaccharide. In another more preferred embodiment, the oligosaccharide is an oligosaccharide comprising one or more fucose groups and one or more sialic acid groups, which is selected from Neu4Ac; Neu5Ac; Neu4,5Ac2; Neu5,7Ac2; Neu5,8Ac2; Neu5,9Ac2; Neu4,5,9Ac3; Neu5,7,9Ac3; Neu5,8,9Ac3; Neu4,5,7,9Ac4; Neu5,7,8,9Ac4; Neu4,5,7,8,9Ac5; Neu5Gc and KDO.

[0062] In the context of the present invention, the solution comprising oligosaccharides to be purified by the method of the present invention is a solution selected from the enumeration of any process stream comprising biocatalytic reaction solution, chemosynthesis solution, cell culture and method as described herein. More specifically, the solution comprising oligosaccharides to be purified by the method of the present invention is a solution selected from the enumeration of biocatalytic reaction solution, chemosynthesis solution, cell culture and fermentation as described herein. In the context of the present invention, the oligosaccharides are produced by the biocatalytic reaction solution, the chemosynthesis solution or by the cells cultivated in the cell culture or by the fermentation. In the context of the present invention, the solution comprising the oligosaccharides is produced by incubating in a reactor as defined herein or in an incubator. The reactor or incubator can vary from small-scale size (laboratory scale) to large-scale size (industrial setting).

[0063] In preferred embodiments, the purity of the oligosaccharide in the solution prior to purification by the method is <70%, <60%, <50%, <40%, <30%, <20%, <10% based on total dry solids.

[0064] In another and / or additional preferred embodiment, the oligosaccharide is accompanied in the solution by sialic acid as defined herein; ash, preferably, the ash comprises sulfate, phosphate, sodium, chloride, potassium and heavy metals, preferably the heavy metals include zinc, manganese, copper, ammonium, lead, arsenic, cadmium and / or mercury; one or more monosaccharides, such as fucose (Fuc), galactose (Gal), glucose (Glc), N-acetylglucosamine (GlcNAc), N-acetylgalactosamine (GalNAc), mannose (Man), N-acetylmannosamine (ManNAc); one or more activated monosaccharides, such as UDP-N-acetylglucosamine (UDP-GlcNAc), UDP-N-acetylgalactosamine (GalNAc), mannose (Man), N-acetylmannosamine (ManNAc); one or more activated monosaccharides, such as UDP-N-acetylglucosamine (UDP-GlcNAc), UDP-N-acetylgalactosamine (GalNAc), mannose (Man), N-acetylmannosamine (ManNAc); one or more activated monosaccharides, such as UDP-N-acetylglucosamine (UDP- Lactosamine (UDP-GalNAc), UDP-N-acetylmannosamine (UDP-ManNAc), UDP-glucose (UDP-Glc), UDP-galactose (UDP-Gal), GDP-mannose (GDP-Man), GDP-fucose (GDP-Fuc), UDP-glucuronate, UDP-galacturonate, CMP-sialic acid (CMP-Neu5Ac), CMP-Neu4Ac, CMP-Neu5Ac9N3, CMP-Neu4,5Ac2, CMP-Neu5,7Ac2, CMP-Neu5,9Ac2, CMP-Neu5,7(8,9) ... Acylneuraminic acid (CMP-Neu5Gc); one or more phosphorylated monosaccharides, such as fructose-6-phosphate, glucose-6-phosphate, glucose-1-phosphate, glucosamine-6-phosphate, N-acetylglucosamine-6-phosphate, N-acetylglucosamine-1-phosphate, galactose-1-phosphate, N-acetylmannosamine-6-phosphate, mannose-6-phosphate, mannose-1-phosphate, fructose-1-phosphate, fructose-1,6-bisphosphate, glycerol-3-phosphate, glyceraldehyde-3-phosphate, dihydroxyacetone-phosphate; one or more disaccharides, such as lactose (Gal-β1,4-Glc), lactose-N-disaccharide (Gal-β1,3-GlcNAc), N-acetyllactosamine (Gal-β1,4-GlcNAc) and / or one or more other oligosaccharides selected from the list including: neutral (uncharged) oligosaccharides, negatively charged oligosaccharides, milk oligosaccharides, preferably mammalian milk oligosaccharides (MMO), more preferably human milk oligosaccharides (HMO); O-antigens; Enterobacterial common antigens (ECA); oligosaccharide repeating units present in capsular polysaccharides; peptidoglycans; amino sugars; Lewis-type antigenic oligosaccharides; antigens of the human ABO blood group system; animal oligosaccharides, preferably selected from N-glycans and O-glycans; plant oligosaccharides, preferably selected from N-glycans and O-glycans; glucopyranosylsucrose (Glc-α1,4-Glc-α1,2-Fru); lactulose-N-triose II (GlcNAc-b1,3-Gal-b1,4-Fru); lactulose-N-tetraose; lactulose-N-neotetraose; uridine diphosphate galactose; preferably selected from the group consisting of the following fucosylated oligosaccharides: 2'-fucosyllactose (2'FL), 3-fucosyllactose (3FL), 4-fucosyllactose (4FL), 6-fucosyllactose (6FL), 2',3-difucosyllactose (diFL), lactose-N-fucopentaose I (LNFP I), Gal-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (Gal-LNFP I), GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (GalNAc-LNFP I), lactose-N-neofucopentaose I (LNnFP I), lactose-N-fucopentaose II (LNFP II), lactose-N-fucopentaose III (LNFP III), lactose-N-fucopentaose V (LNFP V), lactose-N-fucopentaose VI (LNFP VI), lactose-N-neofucopentaose V, lactose-N-difucohexaose I (LNDFH-I), lactose-N-difucohexaose II (LNDFH-II), Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-GlcNAc-b1,3 -Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1 ,4-Glc,Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc,Fuc-a1,2- Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc、Fuc-a1,4-(Fuc-a1,2-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc、monofucosyllactose-N-hexaose-III、difucosyllactose Lactose-hexaose (a), difucosyl-lactose-N-hexaose, difucosyl-lactose-N-neohexaose, trifucosyllactose-N-hexaose, a1,3-galactosyl-3-fucosyllactose, Gal-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, 2-fucosyllactulose, 3-fucosyl-N-acetyllactosamine, 2'-fucosyl-N-acetyllactosamine, difucosyl-N-acetyllactosamine, 4-fucosyllactose-N-disaccharide, 2'-fucosyllactose-N-disaccharide, difucosyllactose-N-disaccharide and GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc; preferably selected from the group consisting of sialylated oligosaccharides including 3' sialyllactose (3'SL), 6' sialyllactose (6'SL), 8' sialyllactose (8'SL), 3,6-disialyllactose (Neu5Ac-α2,3-(Neu5Ac-α2,3-)Glc) and 4-(Fuc-a1,3-)Glc. Ac-α2,6)-Gal-β1,4-Glc), 6,6'-disialyl lactose (Neu5Ac-α2,6-Gal-β1,4-(Neu5Ac-α2,6)-Glc), 8,3-disialyl lactose (Neu5Ac-α2,8-Neu5Ac-α2,3-Gal-β1,4-Glc), 6'-sialyl lactosamine, 3'-sialyl lactosamine, sialyllactose-N-tetraose a (LSTa), sialyllactose-N-tetraose b (LSTb), sialyllactose-N-tetraose c (LSTc), sialyllactose-N-tetraose d (LSTd), disialyl lactose-N-tetraose, disialyl lactose-N-neotetraose , monosialyllactose-N-hexaose, disialyllactose-N-hexaose I, disialyllactose-N-hexaose II, monosialyllactose-N-neohexaose I, monosialyllactose-N-neohexaose II, disialyllactose-N-neohexaose, 3'-sialyl-3-fucosyllactose, fucodisialyllactose-N-hexaose, disialylmonofucosyllactose-N-neohexaose, sialyllactose-N-fucohexaose II, disialyllactose-N-fucopentaose II, monofucosyldisialyllactose-N-tetraose, Neu5Ac-a2,3-Gal-b1,4-GlcNAc-b1,3-Gal, Neu5Ac-a2,3-Gal-b1,3-GlcNAc c-b1,3-Gal, 3'-KDO-lactose, 3'-KDO-lactosamine, 3'-KDO-6' sialyllactose, 3'KDO-8-sialyllactose, KDO-2,3Galβ-1,3GalNacβ-1,3Galα-1,4Galβ-1,4Gal, KDO-2,3Galβ-1,3GlcNacβ-1,3Galβ-1,4Glc, KDO-2,3Galβ-1,4GlcNacβ-1,3Galβ-1,4Glc, 3'-KDO-3-fucosyllactose, Neu5Ac-a2,8-Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-sialyl-2'-fucosyllactose, 6'-sialyl-2'-fucosyllactose, 6'-sialyl-3-fucosyllactose, 3'-sialyl-3-fucosyllactose, Neu5Ac-a2,6-(Neu5Ac-a2,3-)Gal-b1,4-Glc, 3'-sialyl-3-fucosyllactosamine, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc, 6'-sialyl lactose-N-biose, 3'-sialyl lactose-N-biose, Neu5Ac-a2,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, Neu 5Ac-a2,6-(Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,3-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6 -(Neu5Ac-a2,6-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal- b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-Gal-b1,3-GlcN Ac-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-Gal-b1,3-GlcNA c-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc-b1,3-Ga l-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc and any combination thereof; N-acetylglucosamine-containing oligosaccharides, which are preferably selected from the following list: lactose-N-triose II (LN3), lactose-N-tetraose (LNT), lactose-N-neotetraose (LNnT), 6'-galactosyl lactose, 4'-galactosyl lactose, 3'-galactosyl lactose , GlcNAc-b1,6-Gal-b1,4-Glc, lactose-N-hexaose (LNH), lactose-N-neohexaose (LNnH), p-lactose-N-hexaose (pLNH), p-lactose-N-neohexaose (pLNnH), GlcNAc-b1,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, lactose-N-pentaose (LN5), lactose-N-neopentaose, p-lactose-N-pentaose, p-lactose-N-neopentaose, lactose-N-neopentaose I, lactose-N-heptaose (LN7), lactose-N-neoheptaose, p-lactose-N-neoheptaose, p-lactose-N-heptaose, lactose -N-octaose (LNO), lactose-N-neooctaose, allolactose-N-octaose, p-lactose-N-octaose, allolactose-N-neooctaose, novel lactose-N-neooctaose, p-lactose-N-neooctaose (pLNnO), allolactose-N-nonaose, novel lactose-N-nonaose, lactose-N-nonaose (LN9), lactose-N-decaose, allolactose-N-decaose, novel lactose-N-decaose, lactose-N-neodecaose, p-lactose-N-neodecaose (pLNnD), a1,3-galactosyl lactose-N-neotetraose, GlcNAc-b1,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Gl c. GlcNAc-b1,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc and GlcNAc-b1,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc; neutral (uncharged) oligosaccharides containing N-acetylglucosamine; oligosaccharides containing N-acetyllactosamine; oligosaccharides containing lactose-N-disaccharide; non-fucosylated neutral (uncharged) oligosaccharides; chitosan; oligosaccharides containing chitosan; heparin precursor; chondroitin sulfate; glycosaminoglycan oligosaccharides; heparin; heparan sulfate; dermatan sulfate; hyaluronan; hyaluronic acid; and keratan sulfate.

[0065] In another and / or additional preferred embodiment, the solution comprises two or more oligosaccharides.

[0066] In another and / or additional preferred embodiment, the oligosaccharide is accompanied in the solution by one or more other oligosaccharides, wherein at least one of the other oligosaccharides has the same degree of polymerization (DP) as the oligosaccharide. In a more preferred embodiment, the oligosaccharide is accompanied in the solution by one or more other oligosaccharides, wherein all of the other oligosaccharides have the same DP as the oligosaccharide. In an alternative preferred embodiment, the oligosaccharide is accompanied in the solution by one or more other oligosaccharides, wherein at least one of the other oligosaccharides has a different degree of polymerization (DP) than the oligosaccharide. In a more preferred alternative embodiment, the oligosaccharide is accompanied in the solution by one or more other oligosaccharides, wherein all of the other oligosaccharides have a different DP than the oligosaccharide.

[0067] In another preferred embodiment, the solution comprises 6'SL, lactose, and sialic acid. In another preferred embodiment, the solution comprises 3'SL, lactose, and sialic acid. In another preferred embodiment, the solution comprises 3'SL and 6'SL. In another preferred embodiment, the solution comprises sialic acid, lactose, 3'SL, and 6'SL.

[0068] In another preferred embodiment, the solution comprises sialic acid, LSTc, and 6'SL. In another preferred embodiment, the solution comprises sialic acid, lactose, LSTc, and 6'SL. In another preferred embodiment, the solution comprises sialic acid, LN3, LNnT, LSTc, and 6'SL. In another preferred embodiment, the solution comprises sialic acid, lactose, LN3, LNnT, LSTc, and 6'SL. In another preferred embodiment, the solution comprises sialic acid, LN3, sialylated LN3, LNnT, LSTc, and 6'SL. In another preferred embodiment, the solution comprises sialic acid, LN3, sialylated LN3, LNnT, lactose-N-hexaose, LSTc, and 6'SL.

[0069] In another preferred embodiment, the solution comprises sialic acid, LSTa, and 3'SL. In another preferred embodiment, the solution comprises sialic acid, lactose, LSTa, and 3'SL. In another preferred embodiment, the solution comprises sialic acid, LN3, LNT, LSTa, and 3'SL. In another preferred embodiment, the solution comprises sialic acid, lactose, LN3, LNT, LSTa, and 3'SL. In another preferred embodiment, the solution comprises sialic acid, LN3, sialylated LN3, LNT, LSTa, and 3'SL. In another preferred embodiment, the solution comprises sialic acid, LN3, sialylated LN3, LNT, lactose-N-hexaose, LSTa, and 3'SL.

[0070] In another preferred embodiment, the solution comprises 2'FL, 3-FL, 3'SL, and 6'SL. In another preferred embodiment, the solution comprises 2'FL, 3-FL, 3'SL, 6'SL, and lactose. In another preferred embodiment, the solution comprises 2'FL, 3-FL, 3'SL, 6'SL, sialic acid, and lactose. In another preferred embodiment, the solution comprises 2'FL, LNFP-I, 3'SL, and LSTa. In another preferred embodiment, the solution comprises 3-FL, LNFP-III, 6'SL, and LSTc. In another preferred embodiment, the solution comprises 2'FL, 3-FL, di-FL, 3'SL, 6'SL, LNT, and LNnT. In another preferred embodiment, the solution comprises LSTc and LSTa.

[0071] In another preferred embodiment, the solution comprises only one or more neutral (uncharged) fucosylated oligosaccharides as defined herein. In another preferred embodiment, the solution comprises one or more neutral (uncharged) fucosylated oligosaccharides and one or more charged fucosylated oligosaccharides. Charged fucosylated oligosaccharides are understood to be oligosaccharides comprising at least one fucose residue and at least one sialic acid residue as defined herein. In another preferred embodiment, the solution comprises only one or more neutral (uncharged) oligosaccharides. In another preferred embodiment, the solution comprises one or more neutral oligosaccharides and one or more charged oligosaccharides. In another preferred embodiment, the solution comprises one or more neutral (uncharged) fucosylated oligosaccharides and one or more charged oligosaccharides. In another preferred embodiment, the solution comprises one or more neutral (uncharged) fucosylated oligosaccharides and one or more charged oligosaccharides. In another preferred embodiment, the solution comprises one or more neutral (uncharged) fucosylated oligosaccharides and one or more sialylated oligosaccharides.

[0072] In another and / or additional preferred embodiment of the method of the present invention, the ash content of the solution comprising the oligosaccharides to be purified by the method, prior to purification by the method of the present invention, is >10%, based on total dry solids. The ash content of the solution can be determined as described herein.

[0073] In another and / or additional preferred embodiment of the process of the present invention, the solution comprising the oligosaccharides to be purified by the process of the present invention has an ash content of >10% based on total dry solids before purification by said process, wherein said ash comprises any one or more heavy metals selected from the group consisting of lead, arsenic, cadmium, mercury, zinc, manganese, copper, iron, magnesium and calcium.

[0074] In a further and / or additional preferred embodiment of the process according to the invention, the solution comprising oligosaccharides to be purified by the process according to the invention comprises, before purification by said process, a lead content of >0.1 mg / kg dry solids, an arsenic content of >0.2 mg / kg dry solids, a cadmium content of >0.1 mg / kg dry solids and / or a mercury content of >0.5 mg / kg dry solids.

[0075] In another and / or additional preferred embodiment, the solution comprising oligosaccharides to be purified by the method of the present invention is a cell culture using cells producing the oligosaccharides and comprising the oligosaccharides, biomass, culture medium components, additives and pollutants (e.g., defoaming agents). In a more preferred embodiment, the purity of the oligosaccharides in the cell culture is <70%, <60%, <50%, <40%, <30%, <20%, <10% based on total dry solids before purification by the method. In another and / or additional more preferred embodiment, the biomass (when present) separated during the method is optionally recycled to the cell culture.

[0076] In a preferred embodiment, the oligosaccharides purified by the method of the present invention are produced by cells cultured in cell culture. In the context of the present invention, cell culture includes in vitro and / or in vitro culture of cells. In another preferred embodiment of the present invention, the cell is a bacterium, fungus, yeast, plant cell, animal cell or protozoan cell.

[0077] In another and / or extra preferred embodiment, cell is a metabolically engineered cell.In another and / or extra preferred embodiment, cell is not any one or more compounds of oligosaccharide by metabolically engineered to produce.In another and / or extra preferred embodiment, cell is metabolically engineered to produce oligosaccharide.In even more preferred embodiment, cell is metabolically engineered to produce two or more oligosaccharides by metabolically engineered.

[0078] In another and / or additional preferred embodiment, the cell produces oligosaccharides and any one or more sialic acids as defined herein; one or more monosaccharides; one or more activated monosaccharides; one or more phosphorylated monosaccharides; one or more disaccharides and / or one or more other oligosaccharides, as described herein. In another and / or additional preferred embodiment, the cell has been metabolically engineered to produce oligosaccharides and any one or more sialic acids as defined herein; one or more monosaccharides, one or more activated monosaccharides, one or more phosphorylated monosaccharides; one or more disaccharides and / or one or more other oligosaccharides as described herein.

[0079] In another and / or additional preferred embodiment, the solution is a cell culture using at least one cell that has been metabolically engineered to produce the oligosaccharide and one or more of: i) sialic acid, ii) one or more monosaccharides, iii) one or more activated monosaccharides, iv) one or more phosphorylated monosaccharides, v) one or more disaccharides and / or vi) one or more other oligosaccharides.

[0080] In a more preferred embodiment, the cell comprises a sialylation pathway. In an even more preferred embodiment, the cell is metabolically engineered to comprise a sialylation pathway. In another even more preferred embodiment, the cell is metabolically engineered to comprise a sialylation pathway selected from the group consisting of L-glutamine-D-fructose-6-phosphate aminotransferase, phosphoglucosamine mutase, N-acetylglucosamine-6-P deacetylase, N-acylglucosamine 2-epimerase, UDP-N-acetylglucosamine 2-epimerase, N-acetylmannosamine-6-phosphate 2-epimerase, UDP-GlcNAc Any one or more genes listed in the group consisting of d-epimerase / kinase, glucosamine 6-phosphate N-acetyltransferase, N-acetylglucosamine-6-phosphate phosphatase, phosphoacetylglucosamine mutase, N-acetylglucosamine 1-phosphate uridyltransferase, glucosamine-1-phosphate acetyltransferase, Neu5Ac synthase, N-acetylneuraminate lyase, N-acylneuraminate-9-phosphate synthase, N-acylneuraminate-9-phosphatase, d-arabinose 5-phosphate isomerase, KDO-8P synthase, KDO 8-phosphate phosphatase, sialic acid transporter, CMP kinase, CMP-sialic acid synthase, and sialyltransferase have modified and / or enhanced expression.

[0081] In another and / or additional preferred embodiment, the cell comprises a fucosylation pathway. In a more preferred additional and / or alternative embodiment, the cell is metabolically engineered to comprise a fucosylation pathway. In another even more preferred additional and / or alternative embodiment, the cell has been metabolically engineered to comprise a fucosylation pathway, wherein any one or more genes selected from the group consisting of mannose-6-phosphate isomerase, phosphomannose mutase, mannose-1-phosphate guanylyltransferase, GDP-mannose 4,6-dehydratase, GDP-L-fucose synthase, fucose permease, fucokinase, fucose-1-phosphate guanylyltransferase, and fucosyltransferase have modified and / or enhanced expression.

[0082] In another and / or additional preferred embodiment, the cell comprises a galactosylation pathway. In a more preferred additional and / or alternative embodiment, the cell is metabolically engineered to comprise a galactosylation pathway. In another even more preferred additional and / or alternative embodiment, the cell is metabolically engineered to comprise a galactosylation pathway, wherein any one or more genes selected from the group consisting of galactose-1-epimerase, galactokinase, glucokinase, galactose-1-phosphate uridyltransferase, UDP-glucose 4-epimerase, glucose-1-phosphate uridyltransferase, phosphoglucomutase, and galactosyltransferase have modified and / or enhanced expression.

[0083] In another and / or additional preferred embodiment, the cell comprises an "N-acetylglucosaminyl" pathway. In a more preferred further and / or alternative embodiment, the cell is metabolically engineered to comprise an N-acetylglucosaminyl pathway. In another even more preferred further and / or alternative embodiment, the cell has been metabolically engineered to comprise an N-acetylglucosaminyl pathway in which any one or more genes selected from the group consisting of L-glutamine-D-fructose-6-phosphate aminotransferase, N-acetylglucosamine-6-phosphate deacetylase, phosphoglucosamine mutase, N-acetylglucosamine-1-phosphate uridyltransferase, glucosamine-1-phosphate acetyltransferase, and a glycosyltransferase that transfers GlcNAc have modified and / or enhanced expression.

[0084] In another and / or additional preferred embodiment, the cell comprises an "N-acetylgalactosaminyl" pathway. In a more preferred further and / or alternative embodiment, the cell is metabolically engineered to comprise an N-acetylgalactosaminyl pathway. In another even more preferred further and / or alternative embodiment, the cell has been metabolically engineered to comprise an N-acetylgalactosaminyl pathway in which any one or more genes selected from the group consisting of L-glutamine-D-fructose-6-phosphate aminotransferase, phosphoglucosamine mutase, N-acetylglucosamine 1-phosphate uridyltransferase, glucosamine-1-phosphate acetyltransferase, UDP-N-acetylglucosamine 4-epimerase, UDP-glucose 4-epimerase, N-acetylgalactosamine kinase and / or UDP-N-acetylgalactosamine pyrophosphorylase and glycosyltransferases that transfer GalNAc have modified and / or enhanced expression.

[0085] In another and / or additional preferred embodiment, the cell comprises a "mannosylation" pathway. In a more preferred further and / or alternative embodiment, the cell is metabolically engineered to comprise a mannosylation pathway. In another even more preferred further and / or alternative embodiment, the cell has been metabolically engineered to comprise a mannosylation pathway, wherein any one or more genes selected from the group consisting of mannose-6-phosphate isomerase, phosphomannose mutase, and / or mannose-1-phosphate guanylyltransferase and mannosyltransferase have modified and / or enhanced expression.

[0086] In another and / or additional preferred embodiment, the cell comprises an "N-acetylmannosamination" pathway. In more preferred further and / or alternative embodiments, the cell is metabolically engineered to comprise an N-acetylmannosamination pathway. In another even more preferred additional and / or alternative embodiment, the cell has been metabolically engineered to comprise an N-acetylmannosamination pathway, wherein any one or more genes selected from the list of L-glutamine-D-fructose-6-phosphate aminotransferase, glucosamine-6-phosphate deaminase, phosphoglucosamine mutase, N-acetylglucosamine-6-phosphate deacetylase, glucosamine 6-phosphate N-acetyltransferase, N-acetylglucosamine-1-phosphate uridyltransferase, glucosamine-1-phosphate acetyltransferase, glucosamine-1-phosphate acetyltransferase, UDP-GlcNAc 2-epimerase and / or ManNAc kinase and ManNAc-transferring glycosyltransferases have modified and / or enhanced expression.

[0087] In another and / or additional preferred embodiment, the cell is metabolically engineered to enhance oligosaccharide production, enhance uptake of one or more precursors and / or receptors for oligosaccharide synthesis, better oligosaccharide efflux, reduce production of byproducts such as, for example, acids, increase availability of cofactors such as, for example, ATP, NADP, NADPH, and / or better metabolic flux through any of the sialylation, fucosylation, galactosylation, N-acetylglucosaminylation, N-acetylgalactosaminylation, mannosylation and / or N-acetylmannosaminylation pathways present in the cell.

[0088] In another and / or extra preferred embodiment, cell produces described oligosaccharide from one or more internalization precursors as defined herein.Preferably, described precursor is given cell from culture medium or incubation feed.In a more preferred embodiment, cell synthesis participates in one or more precursors of described oligosaccharide production.In another preferred embodiment, the precursor that cell is used to produce described oligosaccharide is converted into described oligosaccharide completely.In another preferred embodiment, the precursor that uses in the described solution for the production of described oligosaccharide is converted into described oligosaccharide completely.

[0089] In another and / or additionally more preferred embodiment, the cell culture is fermentation.

[0090] In an alternative and / or additional more preferred embodiment, the cells are cultured or incubated in a reactor as defined herein.In an alternative and / or additional more preferred embodiment, the cells are cultured or incubated in an incubator as defined herein.

[0091] In another and / or extra preferred embodiments, culture cells in the substratum comprising a carbon source, the carbon source comprises monose, disaccharides, oligosaccharides, polysaccharides, polyols, glycerine, compound medium, and the compound medium comprises molasses, corn steep liquor, peptone, tryptones or yeast extract.Preferably, the carbon source is selected from glucose, N-acetylglucosamine (GlcNAc), glycerine, fructose, sucrose, maltose, lactose, arabinose, malto-oligosaccharide, maltotriose, sorbitol, wood sugar, rhamnose, galactose, mannose, methanol, ethanol, trehalose, starch, cellulose, hemicellulose, molasses, corn steep liquor, high fructose syrup, acetate, citrate, lactate and pyruvate.In a preferred embodiment, substratum is the substratum that chemical composition determines.In an extra preferred embodiment, substratum is the minimum salt substratum that comprises sulfate, phosphate, chloride, ammonium, calcium, magnesium, sodium, potassium, iron, copper, zinc, manganese, cobalt and / or selenium.

[0092] In another and / or additional preferred embodiment, the cells are cultured in a chemically defined medium.

[0093] In another and / or additional preferred embodiment, the cells are cultured in a minimal salts medium comprising sulfate, phosphate, chloride, ammonium, calcium, magnesium, sodium, potassium, iron, copper, zinc, manganese, cobalt and / or selenium.

[0094] In another and / or additional preferred embodiment, the oligosaccharide-containing solution to be purified by the method of the present invention further comprises phosphate, N-cyclohexyl-3-aminopropanesulfonic acid (CAPS), ethylenediaminetetraacetic acid (EDTA), ethylenebis(oxyethylenenitrilo)tetraacetic acid (EGTA), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), bicarbonate, taurine, glycine, glycerol, sorbitol, sulfonic acid, tris(hydroxymethyl)aminomethane (Tris(hydroxymethyl)aminomethane) s), zwitterionic agents, polyaminosaccharides, carboxymethyl chitosan (CM-CS), CM-CS-HCl, CM-CS-hydroacetic acid, N,N-bis-(2-hydroxyethyl)-2-aminoethanesulfonic acid (BES), 3-(N-morpholino)propanesulfonic acid (MOPS), dimethyl sulfoxide (DMSO), 2-(N-morpholino)ethanesulfonic acid (MES), albumin, amino acid residues, sulfate, chloride, ammonium, calcium, magnesium, sodium, potassium, iron, copper, zinc, manganese, cobalt and / or selenium.

[0095] In another and / or additional preferred embodiment, the solution comprising oligosaccharides to be purified by the process of the present invention further comprises a glycosyl donor, such as, for example, a halide, a hemiacetal, a peracetate, a thioglycoside, a 1,2-orthoester, an O-imidate, a thioimidate, a glycosyl fluoride, a glycosyl ester, a glycosyl carbonate, a thiocyanate, a diaziridine, a xanthate, a glycal, a phosphite, a sulfoxide, a sulfone, a selenoglycoside, an alkenylglycoside, a heteroarylglycoside, a glycosyl iodide, a glycosyl phosphate, a glycosyl disulfide, a Te-glycoside, a glycosylsulfonylcarbamate, a 2-(hydroxycarbonyl)benzylglycoside.

[0096] In another and / or additional preferred embodiment, the solution comprising oligosaccharides to be purified by the method of the present invention further comprises a polar reaction solvent, CH2Cl2, ClCH2CH2Cl, toluene, an ether solvent, a nitrile solvent, benzene, o-dichlorobenzene, urea, (thio)urea, K2CO3, tris(2,4,6-trimethoxyphenyl)phosphine (TTMPP), isobutylene oxide, trimethylsilyl trifluoromethanesulfonate (TMSOTf), N-iodosuccinimide (NIS) and / or trifluoromethanesulfonic acid (TfOH).

[0097] In one embodiment of the invention, the solution is passed through a + The first cation exchange of the cation exchange resin in the form of Na + , K + , Ca 2 + Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4+ In a preferred embodiment, the cation exchange resin in the second cation exchange is Na + form.

[0098] In another embodiment, the first cation exchange comprises H + In a preferred embodiment, the cation exchange resin in the first cation exchange is supplied by a supplier in the form of H + In an alternative preferred embodiment, the cation exchange resin in the first cation exchange is not provided in the form of H + form but is provided by the supplier in another form, such as Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + form and, when used in the method, is regenerated to H by methods known to those skilled in the art. + In alternative and / or additional preferred embodiments, the first cation exchange is present in a single vessel (eg column) in a small-scale (laboratory model) or large-scale (industrial scale) device.

[0099] In another embodiment, the second cation exchanger comprises Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3 + 、Al 3+ or NH4 + form of cation exchange resin.

[0100] In a preferred embodiment, the cation exchange resin in the second cation exchange is Na + In a more preferred embodiment, the cation exchange resin in the second cation exchange is supplied by the supplier in the form of Na + In an alternative preferred embodiment, the cation exchange resin in the second cation exchange is not provided in the form of Na + form but is provided by the supplier in another form, such as H + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe3+ 、Al 3+ or NH4 + form and, when used in the method, is regenerated to Na by methods known to those skilled in the art + form.

[0101] In another preferred embodiment, the cation exchange resin in the second cation exchange is K + In a more preferred embodiment, the cation exchange resin in the second cation exchange is supplied by a supplier in the form of K + In an alternative preferred embodiment, the cation exchange resin in the second cation exchange is not provided in the form of K + form but is provided in another form, such as Na + 、H + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + form and, when used in the method, is regenerated to K by methods known to those skilled in the art + form.

[0102] In another preferred embodiment, the cation exchange resin in the second cation exchange is Ca 2+ In a more preferred embodiment, the cation exchange resin in the second cation exchange is supplied by a supplier in the form of Ca 2+ In an alternative preferred embodiment, the cation exchange resin in the second cation exchange is not provided in the form of Ca 2+ form but is provided in another form, such as Na + 、H + , K + Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + form and, when used in the method, is regenerated to Ca by methods known to those skilled in the art 2+ form.

[0103] In another preferred embodiment, the cation exchange resin in the second cation exchange is Mg 2+ In a more preferred embodiment, the cation exchange resin in the second cation exchange is provided by a supplier in the form of Mg 2+In an alternative preferred embodiment, the cation exchange resin in the second cation exchange is not provided in the form of Mg 2+ form but is provided in another form, such as Na + 、H + , K + , Ca 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + form and, when used in the process, is regenerated to Mg by methods known to those skilled in the art 2+ form.

[0104] In another preferred embodiment, the cation exchange resin in the second cation exchange is Fe 2+ In a more preferred embodiment, the cation exchange resin in the second cation exchange is provided by a supplier in the form of Fe 2+ In an alternative preferred embodiment, the cation exchange resin in the second cation exchange is not provided in the form of Fe 2+ form but is provided in another form, such as Na + 、H + , K + , Ca 2+ Mg 2+ 、Fe 3+ 、Al 3+ or NH4 + form and, when used in the process, is regenerated to Fe by methods known to those skilled in the art. 2+ form.

[0105] In another preferred embodiment, the cation exchange resin in the second cation exchange is Fe 3+ In a more preferred embodiment, the cation exchange resin in the second cation exchange is provided by a supplier in the form of Fe 3+ In an alternative preferred embodiment, the cation exchange resin in the second cation exchange is not provided in the form of Fe 3+ form but is provided in another form, such as Na + 、H + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Al 3+ or NH4 + form and, when used in the process, is regenerated to Fe by methods known to those skilled in the art 3+ form.

[0106] In another preferred embodiment, the cation exchange resin in the second cation exchange is Al 3+ In a more preferred embodiment, the cation exchange resin in the second cation exchange is supplied by a supplier in the form of Al 3+ In an alternative preferred embodiment, the cation exchange resin in the second cation exchange is not provided in the form of Al 3+ form but is provided in another form, such as Na + 、H + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ or NH4 + form and, when used in the process, is regenerated to Al by methods known to those skilled in the art. 3+ form.

[0107] In another preferred embodiment, the cation exchange resin in the second cation exchange is NH4 + In a more preferred embodiment, the cation exchange resin in the second cation exchange is provided by the supplier in the form of NH4 + In an alternative preferred embodiment, the cation exchange resin in the second cation exchange is not provided in the form of NH4 + form but is provided in another form, such as Na + 、H + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ or Al 3+ form and, when used in the process, is regenerated to NH4 by methods known to those skilled in the art + form.

[0108] In alternative and / or additional preferred embodiments, the second cation exchange occurs in a single vessel (eg, column) in a small-scale (laboratory model) or large-scale (industrial scale) apparatus.

[0109] In another and / or additional preferred embodiment, the cation exchange resin present in the first cation exchange is selected from the group consisting of weak acid cation (WAC) exchange resins and strong acid cation (SAC) exchange resins. In another and / or additional preferred embodiment, the cation exchange resin present in the second cation exchange is selected from the group consisting of weak acid cation (WAC) exchange resins and strong acid cation (SAC) exchange resins. The terms "WAC" and "SAC" are generally known in the art. Once the ion exchange resins are balanced, strong ion exchange resins will not significantly lose the charge on their matrix, so a wide pH range can be used. Weak ion exchange resins have a more specific pH range within which they will maintain their charge: typically alkaline to approximately neutral pH in the case of WACs.

[0110] In another and / or additional preferred embodiment, the first cation exchange comprises H + WAC exchange resin in the form of a second cation exchanger containing Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + Form of WAC exchange resin.

[0111] In another and / or additional preferred embodiment, the first cation exchange comprises H + form of SAC exchange resin, and the second cation exchange resin contains Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + Form of WAC exchange resin.

[0112] In another and / or additional preferred embodiment, the first cation exchange comprises H + WAC exchange resin in the form of a second cation exchanger containing Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + form of SAC exchange resin.

[0113] In another and / or additional preferred embodiment, the first cation exchange comprises H + The SAC exchange resin in the form of a second cation exchanger comprises Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + form of SAC exchange resin.

[0114] Ion exchange resin can be for example in packed column, as film, provides as the depth filter cartridge of charge modification, or is used as suspension or fluidized material in the liquid to be processed by ion exchange resin.Ion exchange material is generally included in the matrix that provides different fixed functional groups between cationic exchange material and anionic ion exchange material.The example of suitable ion exchange material comprises fiber gel, microcrystalline gel or beaded gel.These can be made by any material that is for example selected from polysaccharide-based material (for example agarose, sepharose, cellulose), silicon dioxide-based material and organic polymer matrix material (for example polyacrylamide, polystyrene), and its derivatization is to carry anion or cationic group.

[0115] In another and / or additional preferred embodiment, the cation exchange resin present in the first cation exchange is a gel-type version, a porous version or a highly porous version. In another and / or additional preferred embodiment, the cation exchange resin present in the second cation exchange is a gel-type version, a porous version or a highly porous version.

[0116] In another and / or additional preferred embodiment, the cation exchange resin present in the first cation exchange has an acrylic-based, methacrylic-based, styrene-based or polystyrene-based matrix. In another and / or additional preferred embodiment, the cation exchange resin present in the second cation exchange has an acrylic-based, methacrylic-based, styrene-based or polystyrene-based matrix.

[0117] In another and / or additional preferred embodiment, the matrix used in the cation exchange resin in the first and / or second cation exchange further comprises divinylbenzene (DVB). In a more preferred embodiment, DVB is cross-linked to the styrene- or polystyrene-based matrix used in the cation exchange resin in the first and / or second cation exchange.

[0118] Examples of cation exchange resins that can be used in the first and / or second cation exchange include Amberlite FPC22 H (Dupont), Amberlite FPC88 H (Dupont), Amberlite FPC23 UPS (Dupont), Amberlite FPC11 Na (Dupont), Amberlite FPC23 UPS H (Dupont), Diaion PK208 (Mitsubishi chemical), Diaion PK212 (Mitsubishi chemical), Diaion PK216 (Mitsubishi chemical), Diaion PK220 (Mitsubishi chemical), Diaion PK228 (Mitsubishi chemical), Lewatit K 2431 (Lanxess), Lewatit K 2620 (Lanxess), Lewatit MDS1368Ca 320 (Lanxess), Lewatit C 249NS (Lanxess), Dowex 88 (Dow chemical) and Dowex single ball C-600B (Dow chemical). If not yet + The cation exchange resin in the first cation exchange needs to be regenerated by a method known to those skilled in the art to obtain H + If the cation exchange resin has not yet been + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + The cation exchange resin in the second cation exchange needs to be regenerated by a method known to those skilled in the art to obtain Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3 + 、Al 3+ or NH4 + form of cation exchange resin.

[0119] In another and / or additional preferred embodiment, the flow rate through the first and / or second cation exchange is at least 0.5 bed volumes per hour (BV / h). In a more preferred embodiment, the flow rate through the first and / or second cation exchange is at least 1 BV / h. In an even more preferred embodiment, the flow rate through the first and / or second cation exchange is at least 1.5 BV / h. In an even more preferred embodiment, the flow rate through the first and / or second cation exchange is at least 2 BV / h. In an even more preferred embodiment, the flow rate through the first and / or second cation exchange is at least 2.5 BV / h. In a most preferred embodiment, the flow rate through the first and / or second cation exchange is at least 3 BV / h.

[0120] In another and / or additional preferred embodiment, the first and / or second cation exchange performed on the solution in the method is performed at a temperature in the range of 0°C to 80°C (including 0°C and 80°C within the range). In a more preferred embodiment, the first and / or second cation exchange is performed at a temperature in the range of 4°C to 60°C (including 4°C and 60°C within the range). In an even more preferred embodiment, the first and / or second cation exchange is performed at a temperature in the range of 4°C to 40°C (including 4°C and 40°C within the range). In an even more preferred embodiment, the first and / or second cation exchange is performed at a temperature in the range of 10°C to 37°C (including 10°C and 37°C within the range). In an even more preferred embodiment, the first and / or second cation exchange is performed at a temperature in the range of 20°C to 30°C (including 20°C and 30°C within the range). In an even more preferred embodiment, the first and / or second cation exchange is performed at a temperature in the range of 20°C to 25°C (including 20°C and 25°C within the stated range). In an even more preferred embodiment, the first and / or second cation exchange is performed at a temperature in the range of 22°C to 24°C (including 22°C and 24°C within the stated range). In a most preferred embodiment, the first and / or second cation exchange is performed at a temperature in the range of 23°C to 24°C (including 23°C and 24°C within the stated range). In another more preferred embodiment, the first and / or second cation exchange is performed at room temperature. In another more preferred embodiment, at a temperature selected from the group consisting of 0°C, 1°C, 2°C, 3°C, 4°C, 5°C, 6°C, 7°C, 8°C, 9°C, 10°C, 11°C, 12°C, 13°C, 14°C, 15°C, 16°C, 17°C, 18°C, 19°C, 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, 41°C, 42°C, ℃, 42 ℃, 43 ℃, 44 ℃, 45 ℃, 46 ℃, 47 ℃, 48 ℃, 49 ℃, 50 ℃, 51 ℃, 52 ℃, 53 ℃, 54 ℃, 55 ℃, 56 ℃, 57 ℃, 58 ℃, 59 ℃, 60 ℃, 61 ℃, 62 ℃, 63 ℃, 64 ℃, 65 ℃, 66 ℃, 67 ℃, 68 ℃, 69 ℃, 70 ℃, 71 ℃, 72 ℃, 73 ℃, 74 ℃, 75 ℃, 76 ℃, 77 ℃, 78 ℃, 79 ℃ and 80 ℃.

[0121] Carrying out the cation exchange step at a specific temperature should be understood as the temperature regulator of the cation exchanger used in the cation exchange step to the specific temperature and / or the temperature regulator of the solution added into the cation exchange as inflow to the specific temperature.The temperature regulation of the cation exchanger can be carried out by the temperature regulation of the environment of for example resin, the jacket around the cation exchanger and / or the environment using the cation exchanger therein. In the context of the present invention, carrying out the first cation exchange step at a specific temperature should be understood as the temperature regulator of the cation exchanger used in the first cation exchange step to the specific temperature and / or the temperature regulator of the solution added into the first cation exchange as inflow to the specific temperature. In addition in the context of the present invention, carrying out the second cation exchange step at a specific temperature should be understood as the temperature regulator of the cation exchanger used in the second cation exchange step to the specific temperature and / or the temperature regulator of the solution added into the second cation exchange as inflow to the specific temperature.

[0122] In another and / or additional preferred embodiment, the first and / or second cation exchange performed on the solution in the method is performed at a pH in the range of 2 to 5, inclusive. In a more preferred embodiment, the first and / or second cation exchange is performed at a pH in the range of 3 to 5, inclusive. In an even more preferred embodiment, the first and / or second cation exchange is performed at a pH in the range of 4 to 5, inclusive. In another more preferred embodiment, the first and / or second cation exchange is performed at a pH selected from the list consisting of about 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, and 5.

[0123] In another and / or additional preferred embodiment, the method further comprises any one or more processes selected from the list comprising, consisting of, or consisting essentially of: homogenization, clarification, clearing, concentration, centrifugation, decantation, dilution, pH adjustment, temperature adjustment, filtration, ultrafiltration, microfiltration, diafiltration, reverse osmosis, electrodialysis, electrodeionization, nanofiltration, dialysis, use of activated carbon or carbon, use of a solvent, use of an alcohol, use of an aqueous alcohol mixture, use of charcoal, tangential flow high performance filtration, tangential flow ultrafiltration, affinity chromatography, ion exchange, ion exchange chromatography, mixed bed ion exchange, hydrophobic interaction chromatography, gel filtration, ligand exchange chromatography, column chromatography, cation exchange adsorption resins, anion exchange adsorption resins, use of an adsorbent material, use of an ion exchange resin, evaporation, vacuum evaporation, wiped film evaporation, falling film evaporation, pasteurization, enzymatic treatment, decolorization, and drying, and wherein any one or more of the processes (a) is carried out in the presence of H + Before the first cation exchange of the cation exchange resin in the form of, and / or any one or more of (b) in the process comprising Na + , K + , Ca 2+ Mg 2+ 、Fe 2 + 、Fe 3+ 、Al 3+ or NH4 + form, preferably Na + It is to be understood herein that none of the processes described herein involve a cation exchange resin containing H + After the first cation exchange of the cation exchange resin in the form of Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + form, preferably Na + In other words, the second cation exchange is performed before the cation exchange resin containing H + The eluate obtained in the first cation exchange of the cation exchange resin in the form of + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + form, preferably Na +In a more preferred embodiment, during the process, the + Before the first cation exchange of the cation exchange resin in the form of and / or in the presence of Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + form, preferably Na + The second cation exchange of the cation exchange resin in the form of cation exchange resin is followed by performing any one or more of the processes more than once.

[0124] In another and / or additional preferred embodiment, the method further comprises any one or more processes selected from the list comprising, consisting of, or consisting essentially of: homogenization, clarification, clearing, concentration, centrifugation, decantation, dilution, pH adjustment in the range of 2 to 5, temperature adjustment, filtration, ultrafiltration, microfiltration, diafiltration, reverse osmosis, nanofiltration, dialysis, use of activated carbon or carbon, use of a solvent in the range of pH 2 to 5, use of an alcohol, use of an aqueous alcohol mixture, use of charcoal, tangential flow high performance filtration, tangential flow ultrafiltration, affinity chromatography, hydrophobic interaction chromatography, gel filtration, ligand exchange chromatography, column chromatography, cation exchange adsorption resins, use of an adsorbent material, evaporation, vacuum evaporation, wiped film evaporation, falling film evaporation, pasteurization, enzymatic treatment, decolorization, drying, and dissolving in a liquid in the range of pH 2 to 5, and wherein any one or more of the processes is carried out in a liquid comprising H + After the first cation exchange of the cation exchange resin in the form of Na + , K + , Ca 2+ Mg 2 + 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + form, preferably Na + Form a cation exchange resin before the second cation exchange.

[0125] Herein, the pH adjustment in the range of 2 to 5 is understood to mean that the pH of the solution is adjusted to a pH in the range of 2 to 5 (including 2 and 5 within this range) before passing the solution through the second cation exchange as described herein. Preferably, the pH of the solution is adjusted to a pH selected from the group consisting of about 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9 and 5 before passing the solution through the second cation exchange as described herein. Use of a solvent having a pH range of 2 to 5 is understood to include use of a solvent having a pH selected from the list consisting of about 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, and 5. Drying and dissolving in a liquid having a pH range of 2 to 5 is understood to mean drying the solution and dissolving in a liquid having a pH selected from the list consisting of about 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9 and 5.

[0126] In a more preferred embodiment, during the regimen, + After the first cation exchange of the cation exchange resin in the form of Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + form, preferably Na + Any one or more of the processes are performed more than once before the second cation exchange of the cation exchange resin in the form of cation exchange resin.

[0127] In another more preferred embodiment, the method comprises i) passing the solution through a liquid comprising H + ii) passing the solution comprising the salt fully or partially converted into acid through a cation exchange resin comprising Na + , K +, Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + The second cation exchange of the cation exchange resin in the form of Na + form, and iii) further purifying the solution to obtain purified oligosaccharides.

[0128] In another more preferred embodiment, the method comprises i) subjecting the solution comprising the oligosaccharide to any one or more of ultrafiltration, microfiltration and / or nanofiltration, ii) passing the solution through a filter comprising H + iii) passing the solution comprising the salt that has been completely or partially converted into an acid through a cation exchange resin comprising Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + The second cation exchange of the cation exchange resin in the form of Na + form, and iv) further purifying the solution to obtain purified oligosaccharides.

[0129] In another more preferred embodiment, the method comprises i) passing the solution through a liquid comprising H + and / or temperature adjustments selected from the group consisting of about 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, and 5, and then iii) passing the solution comprising the salt fully or partially converted to acid through a cation exchange resin comprising Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 +The second cation exchange of the cation exchange resin in the form of Na + form, and iv) further purifying the solution to obtain purified oligosaccharides.

[0130] In another more preferred embodiment, the method comprises i) passing the solution through a liquid comprising H + 4.5, 4.6, 4.7, 4.8, 4.9, and 5, concentrating, diluting, and / or adjusting the temperature; and then iii) passing the solution comprising the salt that is fully or partially converted to the acid through a cation exchange resin containing NaCl. + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + The second cation exchange of the cation exchange resin in the form of Na + form, and iv) further purifying the solution to obtain purified oligosaccharides.

[0131] In another and / or additional preferred embodiment, the temperature of the solution is adjusted to a temperature between 36°C and 65°C, wherein the temperature is within 5°C of the temperature at which the solution exhibits maximum turbidity. The temperature adjustment can be performed at any time during the process. In a more preferred embodiment, the temperature adjustment is combined with a filtration step. In another more preferred embodiment, the temperature of the solution is adjusted to a temperature between 36°C and 60°C. In an even more preferred embodiment, the temperature of the solution is adjusted to a temperature between 40°C and 55°C. In a most preferred embodiment, the temperature of the solution is adjusted to a temperature between 40°C and 45°C. A temperature of 36°C to 65°C is understood to mean a temperature of 36°C, 37°C, 38°C, 39°C, 40°C, 41°C, 42°C, 43°C, 44°C, 45°C, 46°C, 47°C, 48°C, 49°C, 50°C, 51°C, 52°C, 53°C, 54°C, 55°C, 56°C, 57°C, 58°C, 59°C, 60°C, 61°C, 62°C, 63°C, 64°C or 65°C. A temperature of 36°C to 60°C is understood to mean a temperature of 36°C, 37°C, 38°C, 39°C, 40°C, 41°C, 42°C, 43°C, 44°C, 45°C, 46°C, 47°C, 48°C, 49°C, 50°C, 51°C, 52°C, 53°C, 54°C, 55°C, 56°C, 57°C, 58°C, 59°C or 60°C. A temperature of 40°C to 55°C is understood to be a temperature of 40°C, 41°C, 42°C, 43°C, 44°C, 45°C, 46°C, 47°C, 48°C, 49°C, 50°C, 51°C, 52°C, 53°C, 54°C or 55°C. A temperature of 40°C to 45°C is understood to be a temperature of 40°C, 41°C, 42°C, 43°C, 44°C or 45°C.

[0132] In another and / or additional preferred embodiment, the temperature of the solution is adjusted to a temperature between 0°C and 122°C. The temperature adjustment can be performed at any time during the process. A temperature between 0°C and 122°C is understood to mean 0°C, 1°C, 2°C, 3°C, 4°C, 5°C, 6°C, 7°C, 8°C, 9°C, 10°C, 11°C, 12°C, 13°C, 14°C, 15°C, 16°C, 17°C, 18°C, 19°C, 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, 41°C, 42°C, 43°C, 44°C, 45°C, 46°C, 47°C, 48°C, 49°C, 50°C, 51°C, 52°C, 53°C, 54°C, 55°C, 56°C, 57°C, 58°C, 59°C, 60°C, 61°C, 62°C, 63°C, 64°C ℃、32℃、33℃、34℃、35℃、36℃、37℃、38℃、39℃、40℃、41℃、42℃、43℃、44℃、45℃、46℃、47℃、48℃、49℃、50℃、51℃、52℃、53℃、54℃、55℃、56℃、57℃、58℃、59℃、60℃、61℃、62℃、63℃、6 4℃、65℃、66℃、67℃、68℃、69℃、70℃、71℃、72℃、73℃、74℃、75℃、76℃、77℃、78℃、79℃、80℃、81℃、82℃、83℃、84℃、85℃、86℃、87℃、88℃、89℃、90℃、91℃、92℃、93℃、94℃、95℃、96℃ , 97° C., 98° C., 99° C., 100° C., 101° C., 102° C., 103° C., 104° C., 105° C., 106° C., 107° C., 108° C., 109° C., 110° C., 111° C., 112° C., 113° C., 114° C., 115° C., 116° C., 117° C., 118° C., 119° C., 120° C., 121° C., or 122° C. In a more preferred embodiment, the temperature of the solution is adjusted to a temperature between 2° C. and 80° C. The temperature between 2°C and 80°C is understood to mean 2°C, 3°C, 4°C, 5°C, 6°C, 7°C, 8°C, 9°C, 10°C, 11°C, 12°C, 13°C, 14°C, 15°C, 16°C, 17°C, 18°C, 19°C, 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, 41°C, ℃, 41 ℃, 42 ℃, 43 ℃, 44 ℃, 45 ℃, 46 ℃, 47 ℃, 48 ℃, 49 ℃, 50 ℃, 51 ℃, 52 ℃, 53 ℃, 54 ℃, 55 ℃, 56 ℃, 57 ℃, 58 ℃, 59 ℃, 60 ℃, 61 ℃, 62 ℃, 63 ℃, 64 ℃, 65 ℃, 66 ℃, 67 ℃, 68 ℃, 69 ℃, 70 ℃, 71 ℃, 72 ℃, 73 ℃, 74 ℃, 75 ℃, 76 ℃, 77 ℃, 78 ℃, 79 ℃ or 80 ℃. In an even more preferred embodiment, the temperature of the solution is adjusted to a temperature between 4 ℃ and 60 ℃.A temperature of 4°C to 60°C is understood as a temperature of 4°C, 5°C, 6°C, 7°C, 8°C, 9°C, 10°C, 11°C, 12°C, 13°C, 14°C, 15°C, 16°C, 17°C, 18°C, 19°C, 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, 41°C, 42°C, 43°C, 44°C, 45°C, 46°C, 47°C, 48°C, 49°C, 50°C, 51°C, 52°C, 53°C, 54°C, 55°C, 56°C, 57°C, 58°C, 59°C or 60°C. In an even more preferred embodiment, the temperature of the solution is adjusted to a temperature of 10° C. to 55° C. A temperature of 10° C. to 55° C. is understood as a temperature of 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, or 55° C. In an even more preferred embodiment, the temperature of the solution is adjusted to a temperature of 20° C. to 45° C. By a temperature of 20° C. to 45° C. is understood a temperature of 20° C., 21° C., 22° C., 23° C., 24° C., 25° C., 26° C., 27° C., 28° C., 29° C., 30° C., 31° C., 32° C., 33° C., 34° C., 35° C., 36° C., 37° C., 38° C., 39° C., 40° C., 41° C., 42° C., 43° C., 44° C. or 45° C. In an even more preferred embodiment, the temperature of the solution is adjusted to a temperature of 21° C. to 40° C. By a temperature of 21° C. to 40° C. is understood a temperature of 21° C., 22° C., 23° C., 24° C., 25° C., 26° C., 27° C., 28° C., 29° C., 30° C., 31° C., 32° C., 33° C., 34° C., 35° C., 36° C., 37° C., 38° C., 39° C. or 40° C. In an even more preferred embodiment, the temperature of the solution is adjusted to a temperature of 22° C. to 37° C. A temperature of 22° C. to 37° C. is understood as a temperature of 22° C., 23° C., 24° C., 25° C., 26° C., 27° C., 28° C., 29° C., 30° C., 31° C., 32° C., 33° C., 34° C., 35° C., 36° C. or 37° C. In an even more preferred embodiment, the temperature of the solution is adjusted to a temperature of 25° C. to 30° C. A temperature of 25° C. to 30° C. is understood as a temperature of 25° C., 26° C., 27° C., 28° C., 29° C. or 30° C.

[0133] In another and / or additional preferred embodiment of the present invention, the method comprises subjecting the solution comprising oligosaccharides to a first step of ultrafiltration, a second step of nanofiltration and a third step of pH adjustment in the stated order, and then subjecting the pH adjusted solution to a fourth step of ultrafiltration by a filter comprising H + In another and / or additional preferred embodiment, the method further comprises ultrafiltration, nanofiltration and electrodialysis. In another and / or additional preferred embodiment, the method further comprises ultrafiltration, nanofiltration and electrodeionization. In another and / or additional preferred embodiment, the method further comprises ultrafiltration, nanofiltration and electrodeionization. In another and / or additional preferred embodiment, the method further comprises ultrafiltration, nanofiltration, electrodialysis and electrodeionization. In another and / or additional preferred embodiment of the present invention, the method comprises two consecutive nanofiltration steps. In another and / or additional preferred embodiment of the present invention, the method comprises two consecutive electrodialysis steps. In another and / or additional preferred embodiment, the method does not further comprise electrodialysis. In another and / or additional preferred embodiment, the method does not further comprise electrodeionization. In another and / or additional preferred embodiment, the method further comprises a mixed bed ion exchange comprising a cation exchange resin and an anion exchange resin, wherein the cation exchange resin is selected from any form listed below: H + 、Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ NH4 + and wherein the anion exchange resin is selected from the group consisting of OH-, Cl - and SO3 2- In another and / or additional preferred embodiment, the method comprises two mixed bed ion exchanges. In another and / or additional preferred embodiment, the method comprises two consecutive ultrafiltration steps, and wherein the membrane used in the first ultrafiltration step has a higher molecular weight cut-off than the membrane used in the second ultrafiltration step. In a more preferred embodiment, in the case of a mixture comprising H + Prior to said first cation exchange on a cation exchange resin in the form of Λ / Λ / Λ, said solution comprising said oligosaccharides is subjected to said two consecutive ultrafiltration steps.

[0134] In another and / or additional preferred embodiment, the method further comprises clarification, preferably wherein the clarification is performed by any one or more of microfiltration, centrifugation, flocculation or ultrafiltration.

[0135] In another and / or additional preferred embodiment, the method further comprises the use of a cation exchange adsorption resin, an anion exchange adsorption resin and / or the use of an adsorption material.

[0136] In another and / or additional preferred embodiment, the method further comprises drying, preferably wherein the drying is selected from the group consisting of spray drying, freeze drying, spray freeze drying, freeze spray drying, belt drying, conveyor belt drying, vacuum belt drying, vacuum conveyor belt drying, drum drying, roller drying, vacuum drum drying, vacuum roller drying and stirred film drying.

[0137] In another and / or additional preferred embodiment, the method further comprises filtering, preferably wherein the filtering is performed by using a filter aid and / or a flocculant. Preferably, the filter aid is an adsorbent, more preferably the filter aid is activated carbon.

[0138] In another and / or additional preferred embodiment, the method further comprises ultrafiltration, preferably wherein the molecular weight cut-off of the ultrafiltration is equal to or higher than 1 kDa, 2 kDa, 3 kDa, 4 kDa, 5 kDa, 6 kDa, 7 kDa, 8 kDa, 9 kDa, 10 kDa, 11 kDa, 12 kDa, 13 kDa, 14 kDa, 15 kDa.

[0139] In another and / or additional preferred embodiment, the method further comprises nanofiltration, preferably wherein the nanofiltration membrane used in the nanofiltration has a The size exclusion limit, in other words, the nanofiltration has or size exclusion limit.

[0140] In another and / or additional preferred embodiment, the method further comprises diafiltration, preferably wherein said diafiltration is performed on said solution until a conductivity of ≤ 15 mS / cm is reached, preferably ≤ 10 mS / cm, ≤ 5 mS / cm, ≤ 1 mS / cm, ≤ 0.1 mS / cm, ≤ 0.01 mS / cm, ≤ 0.001 mS / cm. In a more preferred embodiment, the method further comprises diafiltration, wherein said diafiltration is performed on said solution until a conductivity of any of 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 mS / cm is reached.

[0141] In another and / or additional preferred embodiment, the method further comprises microfiltration, preferably wherein the pore openings in the membrane used for microfiltration are in the range of 0.1 to 1 μm (micrometer).

[0142] In another and / or additional preferred embodiment, the method further comprises ultrafiltration, preferably wherein the pore openings in the membrane used for ultrafiltration are in the range of 0.01 to 0.1 μm (micrometers).

[0143] In another and / or additional preferred embodiment, the method further comprises nanofiltration, preferably wherein the nanofiltration uses a membrane having pore openings in the range of 0.001 to 0.01 μm (micrometers).

[0144] In another and / or additional preferred embodiment, the method further comprises reverse osmosis, preferably wherein the pore openings in the membrane used for reverse osmosis are in the range of 0.0001 to 0.001 μm (micrometers).

[0145] In another and / or additional preferred embodiment, the method further comprises nanofiltration, preferably wherein the nanofiltration is performed at a pressure in the range of 5 to 20 bar. In other words, the nanofiltration is performed at a pressure of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 bar.

[0146] In another and / or additional preferred embodiment, the method comprises an enzymatic treatment, preferably wherein the enzymatic treatment comprises incubation of the solution with one or more enzymes selected from the group consisting of glycosidase, lactase, β-galactosidase, fucosidase, sialidase, maltase, amylase, hexaminidase, glucuronidase, trehalase and invertase.

[0147] In another and / or additional preferred embodiment, the method further comprises an enzymatic treatment, preferably wherein the enzymatic treatment converts lactose, sucrose, malto-oligosaccharides, maltotriose, sorbitol, trehalose, starch, cellulose, hemicellulose, lignocellulose, molasses, corn syrup and / or high fructose syrup into monosaccharides.

[0148] In another and / or additional preferred embodiment, the method is a batch process.In an alternative and / or additional preferred embodiment, the method is a continuous process.

[0149] In another and / or additional preferred embodiment, the oligosaccharides are in solution with sialic acid and / or ash, and the oligosaccharides are purified from the sialic acid and / or ash by the method of the present invention. In a more preferred embodiment, the oligosaccharides are in solution with sialic acid and / or ash, and the oligosaccharides are purified from the sialic acid and / or ash by the method of the present invention, the method comprising: passing the solution through a precipitate containing H +The first cation exchange of the cation exchange resin in the form of a cation exchange resin, thereby converting the salt in the solution into an acid completely or partially, and passing the solution containing the salt converted into an acid completely or partially through a cation exchange resin comprising Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + The second cation exchange of the cation exchange resin in the form of Na + form.

[0150] In another and / or additional preferred embodiment, the oligosaccharide is accompanied in solution by i) one or more other oligosaccharides and ii) sialic acid and / or ash, and the oligosaccharide and one or more other oligosaccharides are purified from the sialic acid and / or the ash by the method of the present invention. In a more preferred embodiment, the oligosaccharide is accompanied in solution by i) one or more other oligosaccharides and ii) sialic acid and / or ash, and the oligosaccharide and one or more other oligosaccharides are purified from the sialic acid and / or the ash by the method of the present invention, the method comprising: passing the solution through a precipitate containing H + The first cation exchange of the cation exchange resin in the form of a cation exchange resin, thereby converting the salt in the solution into an acid completely or partially, and passing the solution containing the salt converted into an acid completely or partially through a cation exchange resin comprising Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + The second cation exchange of the cation exchange resin in the form of Na + form.

[0151] In another and / or additional preferred embodiment of the present invention, the purity of the oligosaccharides obtained in the purified oligosaccharide solution at the end of the process is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, based on total dry solids. At least 70% is to be understood as 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%. At least 75% is to be understood as 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%. At least 80% is to be understood as 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%. At least 85% is to be understood as 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%. At least 90% is to be understood as 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%. At least 95% is to be understood as 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5% or 100%. At least 97% is to be understood as 97%, 97.5%, 98%, 98.5%, 99%, 99.5% or 100%. At least 98% is to be understood as 98%, 98.5%, 99%, 99.5% or 100%. At least 99% is to be understood as 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 100%.

[0152] In another and / or additional preferred embodiment of the present invention, the purified yield of oligosaccharides obtained in the purified oligosaccharide solution at the end of the process is at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%. At least 60% is to be understood as 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%. At least 65% is to be understood as 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%. At least 70% is to be understood as 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%. At least 75% is to be understood as 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%. At least 80% is to be understood as 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%. At least 85% is to be understood as 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%. At least 90% is to be understood as 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%. At least 95% is to be understood as 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5% or 100%.At least 97% is to be understood as 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, or 100%. At least 98% is to be understood as 98%, 98.5%, 99%, 99.5%, or 100%. At least 99% is to be understood as 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100%.

[0153] In another and / or additional preferred embodiment of the present invention, the purified oligosaccharide solution obtained at the end of the process has an ash content of ≤10% based on total dry solids, preferably ≤9% based on total dry solids, more preferably ≤8% based on total dry solids, even more preferably ≤7% based on total dry solids, even more preferably ≤6% based on total dry solids, even more preferably ≤5% based on total dry solids, even more preferably ≤4% based on total dry solids, even more preferably ≤3% based on total dry solids, even more preferably ≤2% based on total dry solids, even more preferably ≤1% based on total dry solids, and most preferably ≤0.5% based on total dry solids. In a more preferred embodiment, the purified oligosaccharide solution obtained at the end of the process has an ash content of any of 10%, 9%, 8%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2% or 0.1% based on total dry solids.

[0154] In another and / or additional preferred embodiment of the present invention, the purified oligosaccharide solution obtained at the end of the process has an ash content of ≤ 10% based on total dry solids, wherein the ash comprises one or more heavy metals selected from the group consisting of lead, arsenic, cadmium, mercury, zinc, manganese, copper, iron, magnesium and calcium.

[0155] In another and / or additional preferred embodiment of the present invention, the purified oligosaccharide solution obtained at the end of the process has an ash content of ≤10%, based on total dry solids, preferably a lead content of less than 0.1 mg / kg dry solids, an arsenic content of less than 0.2 mg / kg dry solids, a cadmium content of less than 0.1 mg / kg dry solids and / or a mercury content of less than 0.5 mg / kg dry solids.

[0156] In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the process has a lead content of less than 0.1 mg / kg dry solids, more preferably less than 0.05 mg / kg dry solids, even more preferably less than 0.02 mg / kg dry solids, even more preferably less than 0.01 mg / kg dry solids. Preferably, the purified oligosaccharide solution obtained at the end of the process has an ash content of ≤10%, based on total dry solids, and a lead content of less than 0.05 mg / kg dry solids. More preferably, the purified oligosaccharide solution obtained at the end of the process has an ash content of ≤10%, based on total dry solids, and a lead content of less than 0.02 mg / kg dry solids. Even more preferably, the purified oligosaccharide solution obtained at the end of the process has an ash content of ≤10%, based on total dry solids, and a lead content of less than 0.01 mg / kg dry solids.

[0157] In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the process has an arsenic content of less than 0.2 mg / kg dry solids, more preferably less than 0.1 mg / kg dry solids, even more preferably less than 0.05 mg / kg dry solids, even more preferably less than 0.02 mg / kg dry solids. Preferably, the purified oligosaccharide solution obtained at the end of the process has an ash content of ≤10%, based on total dry solids, and an arsenic content of less than 0.1 mg / kg dry solids. More preferably, the purified oligosaccharide solution obtained at the end of the process has an ash content of ≤10%, based on total dry solids, and an arsenic content of less than 0.05 mg / kg dry solids. Even more preferably, the purified oligosaccharide solution obtained at the end of the process has an ash content of ≤10%, based on total dry solids, and an arsenic content of less than 0.02 mg / kg dry solids.

[0158] In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the process has a cadmium content of less than 0.1 mg / kg dry solids, more preferably less than 0.05 mg / kg dry solids, even more preferably less than 0.02 mg / kg dry solids, even more preferably less than 0.01 mg / kg dry solids. Preferably, the purified oligosaccharide solution obtained at the end of the process has an ash content of ≤10%, based on total dry solids, and a cadmium content of less than 0.05 mg / kg dry solids. More preferably, the purified oligosaccharide solution obtained at the end of the process has an ash content of ≤10%, based on total dry solids, and a cadmium content of less than 0.02 mg / kg dry solids. Even more preferably, the purified oligosaccharide solution obtained at the end of the process has an ash content of ≤10%, based on total dry solids, and a cadmium content of less than 0.01 mg / kg dry solids.

[0159] In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the process has a mercury content of less than 0.5 mg / kg dry solids, more preferably less than 0.2 mg / kg dry solids, even more preferably less than 0.1 mg / kg dry solids, even more preferably less than 0.005 mg / kg dry solids. Preferably, the purified oligosaccharide solution obtained at the end of the process has an ash content of ≤10%, based on total dry solids, and a mercury content of less than 0.2 mg / kg dry solids. More preferably, the purified oligosaccharide solution obtained at the end of the process has an ash content of ≤10%, based on total dry solids, and a mercury content of less than 0.1 mg / kg dry solids. Even more preferably, the purified oligosaccharide solution obtained at the end of the process has an ash content of ≤10%, based on total dry solids, and a mercury content of less than 0.005 mg / kg dry solids.

[0160] In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the method is filter sterilized. In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the method is subjected to endotoxin removal. Preferably, endotoxin removal is performed by filtration through a 3 kDa filter, i.e., by filtration with a membrane having a molecular weight cut-off of 3 kDa.

[0161] In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the process has a protein content of less than 100 mg / kg dry solids, a DNA content of less than 10 ng / g dry solids and / or an endotoxin content of less than 10,000 EU / g dry solids. In a more preferred embodiment, the purified oligosaccharide solution obtained at the end of the process has a protein content of equal to or less than 99 mg / kg dry solids, equal to or less than 95 mg / kg dry solids, equal to or less than 90 mg / kg dry solids, equal to or less than 80 mg / kg dry solids, equal to or less than 70 mg / kg dry solids, equal to or less than 60 mg / kg dry solids, equal to or less than 50 mg / kg dry solids, equal to or less than 40 mg / kg dry solids, equal to or less than 30 mg / kg dry solids, equal to or less than 20 mg / kg dry solids or equal to or less than 10 mg / kg dry solids. In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the process has a DNA content of 9 ng / g dry solid or less, 8 ng / g dry solid or less, 7 ng / g dry solid or less, 6 ng / g dry solid or less, 5 ng / g dry solid or less, 4 ng / g dry solid or less, 3 ng / g dry solid or less, 2 ng / g dry solid or less, 1 ng / g dry solid or less, or is free of DNA. In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the process has an endotoxin content of 8000 EU / g dry solid or less, 5000 EU / g dry solid or less, 3000 EU / g dry solid or less, 1000 EU / g dry solid or less, 800 EU / g dry solid or less, 500 EU / g dry solid or less, 300 EU / g dry solid or less than 100 EU / g dry solid. In an even more preferred embodiment, the purified oligosaccharide solution obtained at the end of the method is free of DNA, protein and / or recombinant genetic material. In another more preferred embodiment, the purified oligosaccharide solution obtained at the end of the method is free of recombinant DNA and / or protein derived from a recombinant microorganism.

[0162] In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the process has less than 125 μg / kg 3-MCPD, wherein the 3-MCPD is the sum of 3-monochloropropanediol (3-MCPD) and 3-MCPD fatty acid esters.

[0163] In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the process has less than 0.5 μg / kg epichlorohydrin.

[0164] In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the process has a conductivity of less than 10 mS / cm at 300 g / L of solution.

[0165] In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the method is further concentrated. Concentration can be performed by one or more of nanofiltration, diafiltration, reverse osmosis, evaporation, vacuum evaporation, wiped film evaporation and falling film evaporation. In another and / or additional preferred embodiment, the method further comprises any one or more of nanofiltration, diafiltration, reverse osmosis, evaporation, vacuum evaporation, wiped film evaporation and falling film evaporation, wherein one or more of nanofiltration, diafiltration, reverse osmosis, evaporation, vacuum evaporation, wiped film evaporation and falling film evaporation is performed more than once in the process.

[0166] In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the process is further concentrated to a syrup of at least 20% dry matter. In a more preferred embodiment, the purified oligosaccharide solution obtained at the end of the process is further concentrated to a syrup of at least 30% dry matter. In a more preferred embodiment, the purified oligosaccharide solution obtained at the end of the process is further concentrated to a syrup of at least 40% dry matter.

[0167] In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the process is first subjected to a polishing step before concentration. For this polishing step, an adsorption material such as activated carbon or charcoal, a cation exchange adsorption resin, an anion exchange adsorption resin or a charge-modified depth filter can be used.

[0168] In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the process is further crystallized. In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the process is further dried to a powder. In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the process is further granulated.

[0169] In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the process is further concentrated by a process to an oligosaccharide concentration of >100 g / L, preferably >200 g / L, more preferably >300 g / L, more preferably >400 g / L, more preferably >500 g / L, more preferably >600 g / L, most preferably 300 g / L to 650 g / L. Preferably, the concentration is carried out at a temperature of <80°C, preferably <60°C, more preferably <50°C, more preferably from 20°C to 50°C, even more preferably from 30°C to 45°C. In this context, 20°C to 50°C is understood as 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, 41°C, 42°C, 43°C, 44°C, 45°C, 46°C, 47°C, 48°C, 49°C or 50°C. In this context, 30°C to 45°C is understood as 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, 41°C, 42°C, 43°C, 44°C or 45°C. Preferably, any of the concentration methods is selected from the list comprising using vacuum evaporation or reverse osmosis or nanofiltration.

[0170] In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the process comprises oligosaccharides concentrated to a concentration of >1.5 M and cooled to a temperature of <25°C, more preferably <8°C, to obtain crystalline material of oligosaccharides. A temperature of <25°C is understood as meaning a temperature of 0°C, 1°C, 2°C, 3°C, 4°C, 5°C, 6°C, 7°C, 8°C, 9°C, 10°C, 11°C, 12°C, 13°C, 14°C, 15°C, 16°C, 17°C, 18°C, 19°C, 20°C, 21°C, 22°C, 23°C, 24°C, 25°C or a temperature below 0°C. A temperature of <8°C is understood as meaning a temperature of 0°C, 1°C, 2°C, 3°C, 4°C, 5°C, 6°C, 7°C or 8°C or below 0°C.

[0171] In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the process has a Brix value of about 8 to about 75%, preferably the purified oligosaccharide solution has a Brix value of about 30 to about 65%.

[0172] In another and / or additional preferred embodiment, the purified oligosaccharide solution obtained at the end of the process is dried by any one or more of the following drying steps selected from the group consisting of spray drying, freeze drying, evaporation, precipitation, spray freeze drying, freeze spray drying, belt drying, conveyor belt drying, vacuum belt drying, vacuum conveyor belt drying, drum drying, roller drying, vacuum drum drying, vacuum roller drying and stirred film drying. In a more preferred embodiment, the purified oligosaccharide solution obtained at the end of the process is dried by spray drying, freeze drying or stirred film drying. In another more preferred embodiment, the pH of the purified oligosaccharide solution is in the range of 2 to 7. In other words, the pH of the purified oligosaccharide solution is any one of the following: 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9 or 7. In another even more preferred embodiment, the pH of the purified oligosaccharide solution is in the range of 3 to 6; in other words, the pH of the purified oligosaccharide solution is any one of 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6. In another even more preferred embodiment, the pH of the purified oligosaccharide solution is in the range of 4 to 5; in other words, the pH of the purified oligosaccharide solution is any one of 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or 5. In another more preferred embodiment, the purified oligosaccharide solution obtained at the end of the process is dried by spray drying, in particular at an oligosaccharide solution concentration of 20-60 (w / v), preferably 30-50 (w / v), more preferably 35-45 (w / v), using a nozzle temperature of 110-150° C., preferably 120-140° C., more preferably 125-135° C. and / or an outlet temperature of 60-80° C., preferably 65-70° C.

[0173] According to a second aspect, the present invention provides a purified oligosaccharide solution, purified oligosaccharide or purified oligosaccharide mixture comprising oligosaccharides obtainable by the method described herein, preferably obtained by the method described herein.The purified oligosaccharide solution may comprise one purified oligosaccharide or a purified oligosaccharide mixture.

[0174] In a preferred and / or additional embodiment, the present invention provides a purified oligosaccharide obtainable by the method described herein, preferably obtained by the method described herein, wherein a purified oligosaccharide solution comprising said purified oligosaccharide is dried. In a more preferred embodiment, the present invention provides a purified oligosaccharide obtainable by the method described herein, preferably obtained by the method described herein, wherein a purified oligosaccharide solution comprising said purified oligosaccharide is spray dried. In an alternative more preferred embodiment, the present invention provides a purified oligosaccharide obtainable by the method described herein, preferably obtained by the method described herein, wherein a purified oligosaccharide solution comprising said purified oligosaccharide is dried by a stirred thin film dryer.

[0175] In another and / or additional preferred embodiment, the present invention provides a purified oligosaccharide obtainable by the process described herein, preferably obtained by the process described herein, wherein the purified oligosaccharide solution comprising said purified oligosaccharide is lyophilized.

[0176] In another and / or additional preferred embodiment, the present invention provides a purified oligosaccharide obtainable by the process described herein, preferably obtained by the process described herein, wherein a purified oligosaccharide solution comprising said purified oligosaccharide is crystallized.

[0177] In another and / or additional preferred embodiment, the present invention provides a purified oligosaccharide obtainable by the process described herein, preferably obtained by the process described herein, wherein a purified oligosaccharide solution comprising said purified oligosaccharide is concentrated to a syrup of at least 20% dry matter. In a more preferred embodiment, the present invention provides a purified oligosaccharide obtainable by the process described herein, preferably obtained by the process described herein, wherein a purified oligosaccharide solution comprising said purified oligosaccharide is concentrated to a syrup of at least 30% dry matter. In an even more preferred embodiment, the present invention provides a purified oligosaccharide obtainable by the process described herein, preferably obtained by the process described herein, wherein a purified oligosaccharide solution comprising said purified oligosaccharide is concentrated to a syrup of at least 40% dry matter.

[0178] In another and / or additional preferred embodiment, the present invention provides a purified oligosaccharide mixture comprising oligosaccharides obtainable, preferably obtained, by the process described herein, wherein a purified oligosaccharide solution comprising said purified oligosaccharide mixture comprising oligosaccharides is dried. In a more preferred embodiment, the present invention provides a purified oligosaccharide mixture comprising oligosaccharides obtainable, preferably obtained, by the process described herein, wherein the purified oligosaccharide solution comprising said purified oligosaccharide mixture comprising oligosaccharides is spray dried. In an alternative more preferred embodiment, the present invention provides a purified oligosaccharide mixture comprising oligosaccharides obtainable, preferably obtained, by the process described herein, wherein the purified oligosaccharide solution comprising said purified oligosaccharide mixture comprising oligosaccharides is dried by a stirred thin film dryer.

[0179] In another and / or additional preferred embodiment, the present invention provides a purified oligosaccharide mixture comprising oligosaccharides obtainable by the process described herein, preferably obtained by the process described herein, wherein the purified oligosaccharide solution comprising said purified oligosaccharide mixture comprising oligosaccharides is lyophilized.

[0180] In another and / or additional preferred embodiment, the present invention provides a purified oligosaccharide mixture comprising oligosaccharides obtainable by the process described herein, preferably obtained by the process described herein, wherein a purified oligosaccharide solution comprising said purified oligosaccharide mixture comprising oligosaccharides is crystallized.

[0181] In another and / or additional preferred embodiment, the present invention provides a purified oligosaccharide mixture comprising oligosaccharides obtainable by the process described herein, preferably obtained by the process described herein, wherein a purified oligosaccharide solution comprising said purified oligosaccharide mixture comprising oligosaccharides is concentrated to a syrup of at least 20% dry matter. In a more preferred embodiment, the present invention provides a purified oligosaccharide mixture comprising oligosaccharides obtainable by the process described herein, preferably obtained by the process described herein, wherein a purified oligosaccharide solution comprising said purified oligosaccharide mixture comprising oligosaccharides is concentrated to a syrup of at least 30% dry matter. In an even more preferred embodiment, the present invention provides a purified oligosaccharide mixture comprising oligosaccharides obtainable by the process described herein, preferably obtained by the process described herein, wherein a purified oligosaccharide solution comprising said purified oligosaccharide mixture comprising oligosaccharides is concentrated to a syrup of at least 40% dry matter.

[0182] In another and / or additional preferred embodiment, the present invention provides oligosaccharides purified according to the methods described herein and having an ash content of ≤10% based on total dry solids after the methods. An ash content of ≤10% based on total dry solids is understood to mean 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5% or less than 0.5% ash or less than 0.1% ash based on total dry solids. Preferably, the oligosaccharide is produced by cell culture.

[0183] In another and / or additional preferred embodiment, the present invention provides oligosaccharides purified according to the method described herein and having an ash content of ≤10% on a total dry solids basis, wherein the ash comprises any one or more heavy metals selected from the group consisting of lead, arsenic, cadmium, mercury, zinc, manganese, copper, iron, magnesium and calcium.

[0184] In another and / or additional preferred embodiment, the present invention provides oligosaccharides purified according to the methods described herein and having a lead content of <0.1 mg / kg dry solids. In a more preferred embodiment, the purified oligosaccharide has a lead content of <0.02 mg / kg dry solids. In an even more preferred embodiment, the purified oligosaccharide has a lead content of <0.01 mg / kg dry solids.

[0185] In another and / or additional preferred embodiment, the present invention provides oligosaccharides purified according to the methods described herein and having an arsenic content of <0.2 mg / kg dry solids. In a more preferred embodiment, the purified oligosaccharide has an arsenic content of <0.05 mg / kg dry solids. In an even more preferred embodiment, the purified oligosaccharide has an arsenic content of <0.02 mg / kg dry solids.

[0186] In another and / or additional preferred embodiment, the present invention provides oligosaccharides purified according to the methods described herein and having a cadmium content of <0.1 mg / kg dry solids. In a more preferred embodiment, the purified oligosaccharide has a cadmium content of <0.01 mg / kg dry solids.

[0187] In another and / or additional preferred embodiment, the present invention provides oligosaccharides purified according to the methods described herein and having a mercury content of <0.5 mg / kg dry solids. In a more preferred embodiment, the purified oligosaccharide has a mercury content of <0.1 mg / kg dry solids. In an even more preferred embodiment, the purified oligosaccharide has a mercury content of <0.005 mg / kg dry solids.

[0188] In another and / or additional preferred embodiment, the present invention provides oligosaccharides purified according to the methods described herein and having a protein content of less than 100 mg / kg dry solids, a DNA content of less than 10 ng / g dry solids and / or an endotoxin content of less than 10,000 EU / g dry solids. In a more preferred embodiment, the present invention provides oligosaccharides purified according to the methods described herein and having a protein content of equal to or less than 99 mg / kg dry solids, equal to or less than 95 mg / kg dry solids, equal to or less than 90 mg / kg dry solids, equal to or less than 80 mg / kg dry solids, equal to or less than 70 mg / kg dry solids, equal to or less than 60 mg / kg dry solids, equal to or less than 50 mg / kg dry solids, equal to or less than 40 mg / kg dry solids, equal to or less than 30 mg / kg dry solids, equal to or less than 20 mg / kg dry solids or equal to or less than 10 mg / kg dry solids. In another and / or additional preferred embodiment, the present invention provides oligosaccharides purified according to the methods described herein and having a DNA content of 9 ng / g dry solid or less, 8 ng / g dry solid or less, 7 ng / g dry solid or less, 6 ng / g dry solid or less, 5 ng / g dry solid or less, 4 ng / g dry solid or less, 3 ng / g dry solid or less, 2 ng / g dry solid or less, 1 ng / g dry solid or less, or DNA-free. In another and / or additional preferred embodiment, the present invention provides oligosaccharides purified according to the methods described herein and having an endotoxin content of 8000 EU / g dry solid or less, 5000 EU / g dry solid or less, 3000 EU / g dry solid or less, 1000 EU / g dry solid or less, 800 EU / g dry solid or less, 500 EU / g dry solid or less, 300 EU / g dry solid or less, or 100 EU / g dry solid or less. In an even more preferred embodiment, the present invention provides oligosaccharides purified according to the methods described herein and free of DNA, recombinant DNA, proteins, proteins derived from recombinant microorganisms and / or recombinant genetic material.

[0189] In another and / or additional preferred embodiment, the present invention provides oligosaccharides purified according to the methods described herein and having less than 125 μg / kg of 3-MCPD, wherein the 3-MCPD is the sum of 3-monochloropropanediol (3-MCPD) and 3-MCPD fatty acid esters.

[0190] In another and / or additional preferred embodiment, the present invention provides an oligosaccharide purified according to the methods described herein and having less than 0.5 μg / kg of epichlorohydrin.

[0191] In another and / or additional preferred embodiment, the present invention provides an oligosaccharide purified according to the methods described herein and having a conductivity of less than 10 mS / cm at 300 g / L solution.

[0192] According to a third aspect, the present invention provides spray-dried oligosaccharides or spray-dried oligosaccharide mixtures comprising oligosaccharides, wherein the oligosaccharides or oligosaccharide mixtures are purified according to the method described herein and wherein the spray-dried oligosaccharides or spray-dried oligosaccharide mixtures obtained after the method have an ash content of ≤10% based on total dry solids. An ash content of ≤10% based on total dry solids is understood to mean 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5% or less than 0.5% ash. Preferably, the oligosaccharides or oligosaccharide mixtures are produced by cell culture.

[0193] According to another aspect, the present invention provides a dry powder of a purified oligosaccharide solution obtained by the method as described herein, wherein the dry powder contains ≤15% by weight of water. A dry powder containing ≤15% by weight of water is understood to be a dry powder containing 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1% or 0% by weight of water. In a preferred embodiment, the powder contains ≤10% by weight of water; in other words, the powder contains 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1% or 0% by weight of water. In a more preferred embodiment, the powder contains ≤ 7% water by weight; in other words, the powder contains 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, or 0% water by weight. In a most preferred embodiment, the powder contains ≤ 5% water by weight; in other words, the powder contains 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, or 0% water by weight.

[0194] According to another aspect, the present invention provides a dried powder, preferably a spray-dried powder, of a purified oligosaccharide solution obtained by the process as described herein, wherein the dried powder, preferably the spray-dried powder, has an average particle size of 50 to 250 μm as measured by laser diffraction. In a preferred embodiment, the dried powder, preferably the spray-dried powder, has an average particle size of 95-120 μm as measured by laser diffraction. In a more preferred embodiment, the dried powder, preferably the spray-dried powder, has an average particle size of 110-120 μm as measured by laser diffraction.

[0195] In another and / or additional preferred embodiment, the present invention provides a dry powder of a purified oligosaccharide or a purified oligosaccharide mixture comprising oligosaccharides, wherein the powder, when redissolved in water at a concentration of 10% (mass / volume), provides a solution having a pH of 4 to 7. In other words, the powder, when redissolved in water at a concentration of 10% (mass / volume), provides a solution having a pH of 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9 or 7. In preferred embodiments, the powder, when redissolved in water at a concentration of 10% (mass / volume), provides a solution having a pH of 4 to 6, i.e., a pH of 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, or 6. In more preferred embodiments, the powder, when redissolved in water at a concentration of 10% (mass / volume), provides a solution having a pH of 4 to 5, i.e., a pH of 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or 5. In an even more preferred embodiment, the powder provides a solution having a pH of 5 to 6, i.e., a pH of 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9 or 6, when redissolved in water at a concentration of 10% (mass / volume).

[0196] In another and / or additional preferred embodiment, the present invention provides a dry powder of a purified oligosaccharide or a purified oligosaccharide mixture comprising oligosaccharides, wherein the powder has an ash content of ≤10%, ≤9%, ≤8%, ≤7%, ≤6%, ≤5%, ≤4%, ≤3%, ≤2%, ≤1% or ≤0.5% based on total dry solids.

[0197] In another and / or additional preferred embodiment, the present invention provides a dry powder of a purified oligosaccharide or a purified oligosaccharide mixture comprising oligosaccharides, wherein the powder has an ash content of ≤10% based on total dry solids, and preferably has a lead content of less than 0.1 mg / kg dry solids, an arsenic content of less than 0.2 mg / kg dry solids, a cadmium content of less than 0.1 mg / kg dry solids and / or a mercury content of less than 0.5 mg / kg dry solids.

[0198] In another and / or additional preferred embodiment, the present invention provides a dry powder of a purified oligosaccharide or a purified oligosaccharide mixture comprising oligosaccharides, wherein the powder has an ash content of ≤10% based on total dry solids and a lead content of less than 0.1 mg / kg dry solids, more preferably a lead content of less than 0.05 mg / kg dry solids, even more preferably a lead content of less than 0.02 mg / kg dry solids, and most preferably a lead content of less than 0.01 mg / kg dry solids.

[0199] In another and / or additional preferred embodiment, the present invention provides a dry powder of a purified oligosaccharide or a purified oligosaccharide mixture comprising oligosaccharides, wherein the powder has an ash content of ≤10% based on total dry solids and an arsenic content of less than 0.2 mg / kg dry solids, more preferably an arsenic content of less than 0.1 mg / kg dry solids, even more preferably an arsenic content of less than 0.05 mg / kg dry solids, and most preferably an arsenic content of less than 0.02 mg / kg dry solids.

[0200] In another and / or additional preferred embodiment, the present invention provides a dry powder of a purified oligosaccharide or a purified oligosaccharide mixture comprising oligosaccharides, wherein the powder has an ash content of ≤10% based on total dry solids and a cadmium content of less than 0.1 mg / kg dry solids, more preferably a cadmium content of less than 0.05 mg / kg dry solids, even more preferably a cadmium content of less than 0.02 mg / kg dry solids, and most preferably a cadmium content of less than 0.01 mg / kg dry solids.

[0201] In another and / or additional preferred embodiment, the present invention provides a dry powder of a purified oligosaccharide or a purified oligosaccharide mixture comprising oligosaccharides, wherein the powder has an ash content of ≤10% based on total dry solids and a mercury content of less than 0.5 mg / kg dry solids, more preferably less than 0.2 mg / kg dry solids, even more preferably less than 0.1 mg / kg dry solids, and most preferably less than 0.005 mg / kg dry solids.

[0202] In another and / or additional preferred embodiment, the present invention provides a dry powder of a purified oligosaccharide or a purified oligosaccharide mixture comprising the oligosaccharide, wherein the powder has a protein content of less than 100 mg / kg dry solid, a DNA content of less than 10 ng / g dry solid and / or an endotoxin content of less than 10,000 EU / g dry solid. In a more preferred embodiment, the present invention provides a dry powder of a purified oligosaccharide or a purified oligosaccharide mixture comprising the oligosaccharide, wherein the powder has a protein content of equal to or less than 99 mg / kg dry solid, equal to or less than 95 mg / kg dry solid, equal to or less than 90 mg / kg dry solid, equal to or less than 80 mg / kg dry solid, equal to or less than 70 mg / kg dry solid, equal to or less than 60 mg / kg dry solid, equal to or less than 50 mg / kg dry solid, equal to or less than 40 mg / kg dry solid, equal to or less than 30 mg / kg dry solid, equal to or less than 20 mg / kg dry solid or equal to or less than 10 mg / kg dry solid. In another and / or additional preferred embodiment, the present invention provides a dry powder of a purified oligosaccharide or a purified oligosaccharide mixture comprising oligosaccharides, wherein the powder has a DNA content of equal to or less than 9 ng / g dry solid, equal to or less than 8 ng / g dry solid, equal to or less than 7 ng / g dry solid, equal to or less than 6 ng / g dry solid, equal to or less than 5 ng / g dry solid, equal to or less than 4 ng / g dry solid, equal to or less than 3 ng / g dry solid, equal to or less than 2 ng / g dry solid, equal to or less than 1 ng / g dry solid, or contains no DNA. In another and / or additional preferred embodiment, the present invention provides a dry powder of a purified oligosaccharide or a purified oligosaccharide mixture comprising oligosaccharides, wherein the powder has an endotoxin content of equal to or less than 8000 EU / g dry solid, equal to or less than 5000 EU / g dry solid, equal to or less than 3000 EU / g dry solid, equal to or less than 1000 EU / g dry solid, equal to or less than 800 EU / g dry solid, equal to or less than 500 EU / g dry solid, equal to or less than 300 EU / g dry solid or equal to or less than 100 EU / g dry solid.

[0203] In another and / or additional preferred embodiment, the present invention provides a dry powder of a purified oligosaccharide or a purified oligosaccharide mixture comprising oligosaccharides, wherein the powder is free of DNA, recombinant DNA, proteins, proteins derived from recombinant microorganisms and / or recombinant genetic material.

[0204] In another and / or additional preferred embodiment, the present invention provides a dry powder of a purified oligosaccharide or a purified oligosaccharide mixture comprising oligosaccharides, wherein the powder has less than 125 μg / kg 3-MCPD, wherein the 3-MCPD is the sum of 3-monochloropropanediol (3-MCPD) and 3-MCPD fatty acid esters.

[0205] In another and / or additional preferred embodiment, the present invention provides a dry powder of a purified oligosaccharide or a purified oligosaccharide mixture comprising oligosaccharides, wherein the powder has less than 0.5 μg / kg epichlorohydrin.

[0206] In another and / or additional preferred embodiment, the present invention provides a dry powder of a purified oligosaccharide or a purified oligosaccharide mixture comprising oligosaccharides, wherein the powder is a spray-dried powder.

[0207] In another and / or additional preferred embodiment, the present invention provides purified oligosaccharides as described herein, wherein any one or more of the purified oligosaccharides are milk oligosaccharides. In another embodiment, the present invention provides a purified oligosaccharide mixture comprising oligosaccharides as described herein, wherein the purified oligosaccharide mixture comprises milk oligosaccharides. In a preferred embodiment, milk oligosaccharides are mammalian milk oligosaccharides (MMO). In a more preferred embodiment, milk oligosaccharides are human milk oligosaccharides (HMO). In another preferred embodiment, milk oligosaccharides are neutral (uncharged) milk oligosaccharides, preferably neutral (uncharged) human milk oligosaccharides (HMO), which are selected from the following enumerations: 2'-fucosyllactose, 3-fucosyllactose, 4-fucosyllactose (4FL), 6-fucosyllactose (6FL), 2', 3-difucosyllactose, lactose-N-triose II, lactose-N-tetraose, lactose-N-neotetraose, lactose-N-fucopentaose I, lactose-N-neofucosyllactose Pentose, lactose-N-fucopentaose II, lactose-N-fucopentaose III, lactose-N-fucopentaose V, lactose-N-neofucopentaose V, lactose-N-fucopentaose VI, lactose-N-difucohexaose I, lactose-N-difucohexaose II, lactose-N-hexaose, lactose-N-neohexaose, p-lactose-N-hexaose, p-lactose-N-neohexaose, 6'-galactosyl lactose, 3'-galactosyl lactose, lactose-N-hexaose and lactose-N-neohexaose and any combination thereof. In an alternative preferred embodiment, the milk oligosaccharide is a sialylated milk oligosaccharide, preferably a sialylated human milk oligosaccharide (HMO), selected from the list comprising: 3' sialyllactose, 6' sialyllactose, sialyllactose-N-tetraose a, sialyllactose-N-tetraose b, sialyllactose-N-tetraose c, sialyllactose-N-tetraose d, disialyllactose-N-tetraose, disialyllactose-N-neotetraose, monosialyllactose-N-hexaose, disialyllactose-N-hexaose, Saccharide I, disialyl lactose-N-hexaose II, monosialyllactose-N-neohexaose I, monosialyllactose-N-neohexaose II, disialyl lactose-N-neohexaose, 3'-sialyl-3-fucosyllactose, fucodisialyl lactose-N-hexaose, disialyl monofucosyllactose-N-neohexaose, sialyl lactose-N-fucohexaose II, disialyl lactose-N-fucopentaose II and monofucosyl disialyl lactose-N-tetraose and any combination thereof.

[0208] In another and / or additional preferred embodiment, the present invention provides a purified oligosaccharide or purified oligosaccharide mixture as described herein, wherein the purified oligosaccharide or purified oligosaccharide mixture a) has a conductivity of less than 10 mS / cm at 300 g / L solution; b) is free of recombinant DNA material, optionally free of any DNA; and / or c) is free of proteins derived from recombinant microorganisms, optionally free of any proteins.

[0209] According to another aspect, the present invention provides purified oligosaccharides comprising an ash content of ≤10% based on total dry solids, wherein the ash comprises any one or more heavy metals selected from the group consisting of lead, arsenic, cadmium, mercury, zinc, manganese, copper, iron, magnesium and calcium.

[0210] In a preferred embodiment, the purified oligosaccharide comprises an ash content of ≤10%, based on total dry solids, wherein said ash content of ≤10%, based on total dry solids, comprises a lead content of less than 0.1 mg / kg dry solids, preferably a lead content of less than 0.02 mg / kg dry solids, more preferably a lead content of less than 0.01 mg / kg dry solids, an arsenic content of less than 0.2 mg / kg dry solids, preferably an arsenic content of less than 0.05 mg / kg dry solids, more preferably an arsenic content of less than 0.02 mg / kg dry solids, a cadmium content of less than 0.1 mg / kg dry solids, preferably a cadmium content of less than 0.01 mg / kg dry solids and / or a mercury content of less than 0.5 mg / kg dry solids, preferably a mercury content of less than 0.1 mg / kg dry solids, more preferably a mercury content of less than 0.005 mg / kg dry solids.

[0211] In another and / or additional preferred embodiment, the purified oligosaccharide is selected from the list comprising: fucosylated oligosaccharides, neutral (uncharged) oligosaccharides, negatively charged oligosaccharides, sialylated oligosaccharides, Lewis-type antigens, oligosaccharides containing N-acetylglucosamine, neutral (uncharged) oligosaccharides containing N-acetylglucosamine, oligosaccharides containing N-acetyllactosamine, oligosaccharides containing lactose-N-diose, oligosaccharides containing galactose, non-fucosylated neutral (uncharged) oligosaccharides, chitosan, oligosaccharides containing chitosan, heparin precursors, chondroitin sulfate, glycosaminoglycan oligosaccharides, heparin , heparan sulfate, dermatan sulfate, hyaluronan or hyaluronic acid, keratan sulfate, milk oligosaccharides, O-antigen, enterobacterial common antigen (ECA), oligosaccharide repeating units present in capsular polysaccharides, peptidoglycan, amino sugars, antigens of the human ABO blood group system, animal oligosaccharides, plant oligosaccharides, glucopyranosylsucrose (Glc-a1,4-Glc-a1,2-Fru), lactulose-N-trisaccharide II (GlcNAc-b1,3-Gal-b1,4-Fru), lactulose-N-tetraose, lactulose-N-neotetraose and uridine diphosphate galactose. In a more preferred embodiment, the fucosylated oligosaccharide is selected from the list comprising 2'-fucosyllactose (2'FL), 3-fucosyllactose (3FL), 4-fucosyllactose (4FL), 6-fucosyllactose (6FL), 2',3-difucosyllactose (diFL), lactose-N-fucopentaose I (LNFPI), Gal-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (Gal-LNFP I), GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (GalNAc-LNFP I), lactose-N-neofucopentaose I (LNnFP I), lactose-N-fucopentaose II (LNFP II), and lactose-N-fucopentaose II (LNFP II). II), lactose-N-fucopentaose III (LNFP III), lactose-N-fucopentaose V (LNFP V), lactose-N-fucopentaose VI (LNFP VI), lactose-N-neofucopentaose V, lactose-N-difucohexaose I (LNDFH-I), lactose-N-difucohexaose II (LNDFH-II), Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc、Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc、Fuc-a1,4-(Fuc-a1,2-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc、monofucosyllactose-N-hexaose-III、difucosyllactose-N-hexaose(a), difucosyllactose-N-hexaose, difucosyllactose-N-neohexaose, trifucosyllactose-N-hexaose, a1,3-galacto Glycosyl-3-fucosyllactose, Gal-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, 2-fucosyllactulose, 3-fucosyl-N-acetyllactosamine, 2'-fucosyl-N-acetyllactosamine, difucosyl-N-acetyllactosamine, 4-fucosyllactose-N-disaccharide, 2'-fucosyllactose-N-disaccharide, difucosyllactose-N-disaccharide, and GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc. In a more preferred embodiment, the sialylated oligosaccharide is selected from 3'-sialyllactose (3'SL), 6'-sialyllactose (6'SL), 8'-sialyllactose (8'SL), 3,6-disialyllactose (Neu5Ac-α2,3-(Neu5Ac-α2,6)-Gal-β1,4-Glc), 6,6'-disialyllactose (Neu5Ac-α2,6-Gal-β1,4-(Neu5Ac-α2,6)-Glc), 8,3-disialyllactose (Neu5Ac-α2,8-Neu5Ac-α2,3-Gal-β1,4-Glc), 6'-sialyllactosamine, 3'-sialyllactosamine, sialyllactose-N-tetrasaccharide a (LSTa), sialyllactose-N- Tetrasaccharide b (LSTb), sialyllactose-N-tetrasaccharide c (LSTc), sialyllactose-N-tetrasaccharide d (LSTd), disialyllactose-N-tetrasaccharide, disialyllactose-N-neotetrasaccharide, monosialyllactose-N-hexasaccharide, disialyllactose-N-hexasaccharide I, disialyllactose-N-hexasaccharide II, monosialyllactose-N-neohexacharide I, monosialyllactose Saccharide-N-neohexaose II, disialyl lactose-N-neohexaose, 3'-sialyl-3-fucosyllactose, fucodisialyl lactose-N-hexaose, disialyl monofucosyllactose-N-neohexaose, sialyl lactose-N-fucohexaose II, disialyl lactose-N-fucopentaose II, monofucosyl disialyl lactose-N-tetraose, Neu5Ac-a2,3-Gal-b1,4-GlcNAc-b1,3-Gal, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-KDO-lactose, 3'-KDO-lactosamine, 3'-KDO-6'-sialyllactose, 3'KDO-8-sialyllactose, KDO-2,3Galβ-1,3GalNacβ-1,3Galα-1,4Galβ-1,4Gal, KDO-2,3Galβ-1,3GlcNacβ-1,3Galβ-1,4Glc, KDO-2,3Galβ-1,4GlcNacβ-1,3Galβ-1,4Glc, 3'-KDO-3-fucose Neu5Ac-a2,8-Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-sialyl-2'-fucosyllactose, 6'-sialyl-2'-fucosyllactose, 6'-sialyl-3-fucosyllactose, 3'-sialyl-3-fucosyllactose, Neu5Ac-a2,6-(Neu5Ac-a2,3-)Gal-b1,4-Glc, 3'-sialyl-3-fucosyllactosamine, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc, 6'-sialyl-N-disaccharide, 3'-sialyl-N-disaccharide, N eu5Ac-a2,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,3-Gal-b1,4-(Fuc-a 1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-G lcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1, 4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1 ,3-Gal-b1,4-Glc, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1, ,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,3-)Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, and any combination thereof. In another and / or additionally more preferred embodiment, the neutral (uncharged) oligosaccharide containing N-acetylglucosamine is selected from lacto-N-triose II (LN3), lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), 6'-galactosyl lactose, 4'-galactosyl lactose, 3'-galactosyl lactose, GlcNAc-b1,6-Gal-b1,4-Glc, lacto- N-hexaose (LNH), lactose-N-neohexose (LNnH), p-lactose-N-hexaose (pLNH), p-lactose-N-neohexose (pLNnH), GlcNAc-b1,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, lactose-N-pentaose (LN5), lactose-N-neopentaose, p-lactose-N-pentaose, p-lactose-N-neopentaose, lactose-N - Novel pentasaccharide I, lactose-N-heptasaccharide (LN7), lactose-N-neoheptasaccharide, p-lactose-N-neoheptasaccharide, p-lactose-N-heptasaccharide, lactose-N-octaose (LNO), lactose-N-neooctaose, allolactose-N-octaose, p-lactose-N-octaose, allolactose-N-neooctaose, novel lactose-N-neooctaose, p-lactose-N-neooctaose (pLNnO), allolactose-N-nonasaccharide, novel lactose-N -nonasaccharide, lactose-N-nonasaccharide (LN9), lactose-N-decaose, allolactose-N-decaose, novel lactose-N-decaose, lactose-N-neodecaose, para-lactose-N-neodecaose (pLNnD), a1,3-galactosyl lactose-N-neotetraose, GlcNAc-b1,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, GlcNAc-b1,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc and GlcNAc-b1,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc. In another and / or additional more preferred embodiment, the milk oligosaccharide is a mammalian milk oligosaccharide (MMO), more preferably a human milk oligosaccharide (HMO). In another and / or additional more preferred embodiment, the animal oligosaccharide is selected from N-glycans and O-glycans. In another and / or additional more preferred embodiment, the plant oligosaccharide is selected from N-glycans and O-glycans.

[0212] To identify the oligosaccharides as described herein, standard methods known in the art, such as methylation analysis, reductive cleavage, hydrolysis, GC-MS (gas chromatography-mass spectrometry), MALDI-MS (matrix-assisted laser desorption / ionization-mass spectrometry), ESI-MS (electrospray ionization-mass spectrometry), HPLC (high performance liquid chromatography with ultraviolet or refractive index detection), HPAEC-PAD (high performance anion exchange chromatography with pulsed current detection), CE (capillary electrophoresis), IR (infrared) / Raman spectroscopy and NMR (nuclear magnetic resonance) spectroscopy can be used to identify monomer building blocks (e.g., monosaccharide or polysaccharide unit composition), the anomeric configuration of the side chains, the presence and position of substituents, degree of polymerization / molecular weight and connection mode. Crystal structures can be analyzed using, for example, solid-state NMR, FT-IR (Fourier transform infrared spectroscopy) and WAXS (wide angle X-ray scattering). The degree of polymerization (DP), DP distribution and polydispersity can be determined, for example, by viscometry and SEC (SEC-HPLC, high performance size exclusion chromatography). In order to identify the monomer components of the oligosaccharide, methods such as acid-catalyzed hydrolysis, HPLC (high performance liquid chromatography) or GLC (gas-liquid chromatography) (after conversion to sugar alcohol acetate) can be used. In order to determine the glycosidic bond, the oligosaccharide is methylated in DMSO with methyl iodide and a strong base, hydrolyzed, reduced to partially methylated sugar alcohol, acetylated to methylated sugar alcohol acetate, and analyzed by GLC / MS (gas-liquid chromatography coupled with mass spectrometry). In order to determine the polysaccharide sequence, partial depolymerization is performed using acid or enzyme to determine the structure. In order to identify the anomeric configuration, the oligosaccharide is subjected to enzymatic analysis, for example, by contacting it with an enzyme specific for a particular type of bond (e.g., β-galactosidase or α-glucosidase, etc.), and NMR can be used to analyze the product.

[0213] In another aspect, the present invention provides a purified oligosaccharide or a purified oligosaccharide mixture as described herein for use in medicine, preferably for use in the prevention or treatment of gastrointestinal disorders.

[0214] In another aspect, the present invention provides the purposes of the purified oligosaccharides obtained by the methods described herein in food or feed preparations, in dietary supplements, in cosmetic ingredients or in pharmaceutical ingredients. In some embodiments, the oligosaccharides are mixed with one or more ingredients suitable for food, feed, dietary supplements, pharmaceutical ingredients, cosmetic ingredients or medicines. The purified oligosaccharides can be used to prepare preparations, as food additives, prebiotics, symbiotic bacteria, for the supplementation of baby food, adult food, baby animal feed, adult animal feed, or as treatment or pharmaceutically active compounds or for cosmetic applications. In another aspect, the present invention provides milk oligosaccharides as described herein as additives in food, preferably as additives in human food and / or pet food, more preferably as the purposes of additives in human baby food. In the context of the present invention, food is human food, preferably baby food, human baby food and / or infant formula or infant supplement, and feed is pet food, animal milk replacer, veterinary product, veterinary feed supplement, nutritional supplement, weaning feed or creep feed.

[0215] In another preferred embodiment, a preparation further comprising at least one prebiotic microorganism is provided. In another preferred embodiment of the present invention, the preparation is a nutritional composition. In a more preferred embodiment, the preparation is a pharmaceutical preparation, a dietary supplement, a milk drink or an infant formula. "Prebiotics" are substances that promote the growth of microorganisms that are beneficial to the host (particularly microorganisms in the gastrointestinal tract). In some embodiments, dietary supplements provide a variety of prebiotics, which include the oligosaccharides, which are prebiotics purified by the methods disclosed in this specification to promote the growth of one or more beneficial microorganisms. Examples of prebiotic ingredients for dietary supplements include other prebiotic molecules (such as HMOs) and plant polysaccharides (such as inulin, pectin, β-glucans and oligoxylose). "Prebiotic" products typically contain live microorganisms that replace or add to the gastrointestinal microflora to benefit the recipient. Examples of such microorganisms include Lactobacillus species (e.g., Lactobacillus acidophilus and Lactobacillus bulgaricus), Bifidobacterium species (e.g., B. animalis, B. longum, and B. infantis (e.g., Bi-26), and Saccharomyces boulardii. In some embodiments, the oligosaccharides produced and / or purified by the methods of the present specification are orally administered in combination with such microorganisms. Examples of further ingredients for dietary supplements include oligosaccharides (such as 2'-fucosyllactose, 3-fucosyllactose, 3'-sialyllactose, 6'-sialyllactose), disaccharides (such as lactose), monosaccharides (such as glucose, galactose, L-fucose, sialic acid, glucosamine and N-acetylglucosamine), thickeners (such as gum arabic), acidity regulators (such as trisodium citrate), water, skim milk and flavorings.

[0216] In some embodiments, the oligosaccharide purified by the method as described herein is incorporated into human infant food (e.g., infant formula). Infant formula is typically a manufactured food for feeding infants as a complete or partial substitute for human breast milk. In some embodiments, infant formula is sold as a powder and is prepared by mixing with water for bottle feeding or cup feeding of infants. The composition of infant formula is typically designed to roughly simulate human breast milk. In some embodiments, the oligosaccharide purified by the method as described herein is included in infant formula, to provide a nutritional benefit similar to the nutritional benefit provided by the oligosaccharides in human breast milk. In some embodiments, the purified oligosaccharide is mixed with one or more ingredients of infant formula. Examples of infant formula ingredients include skim milk, a carbohydrate source (e.g., lactose), a protein source (e.g., whey protein concentrate and casein), a fat source (e.g., vegetable oils—such as palm oil, high oleic safflower oil, rapeseed oil, coconut oil, and / or sunflower oil, and fish oil), vitamins (e.g., vitamins A, Bb, B12, C, and D), minerals (e.g., potassium citrate, calcium citrate, magnesium chloride, sodium chloride, sodium citrate, and calcium phosphate), and possibly human milk oligosaccharides (HMOs). In some embodiments, one or more infant formula ingredients include skim milk, a carbohydrate source, a protein source, a fat source, and / or vitamins and minerals. In some embodiments, one or more infant formula ingredients include lactose, whey protein concentrate, and / or high oleic safflower oil. In some embodiments, the concentration of oligosaccharides in the infant formula is about the same as the concentration of oligosaccharides typically found in human breast milk. In some embodiments, oligosaccharides purified by the methods described herein are added to the infant formula at a concentration that is about the same as the concentration of compounds typically found in human breast milk.

[0217] In another aspect, the present invention provides a sugar composition comprising an ash content of ≤10% based on total dry solids and an oligosaccharide content of ≥90% based on total dry solids, wherein the ash comprises any one or more heavy metals selected from the group consisting of lead, arsenic, cadmium, mercury, zinc, manganese, copper, iron, magnesium and calcium. Preferably, the ash content of ≤10% based on total dry solids comprises a lead content of less than 0.1 mg / kg dry solids, preferably a lead content of less than 0.02 mg / kg dry solids, more preferably a lead content of less than 0.01 mg / kg dry solids, an arsenic content of less than 0.2 mg / kg dry solids, preferably a arsenic content of less than 0.05 mg / kg dry solids, more preferably a arsenic content of less than 0.02 mg / kg dry solids, a cadmium content of less than 0.1 mg / kg dry solids, preferably a cadmium content of less than 0.01 mg / kg dry solids and / or a mercury content of less than 0.5 mg / kg dry solids, preferably a mercury content of less than 0.1 mg / kg dry solids, more preferably a mercury content of less than 0.005 mg / kg dry solids. In a more preferred embodiment, the sugar composition comprises an oligosaccharide content of ≥90% based on total dry solids and a lead content of less than 0.1 mg / kg dry solids, preferably less than 0.02 mg / kg dry solids, more preferably less than 0.01 mg / kg dry solids.

[0218] In another and / or additional preferred embodiment, the sugar composition comprises more than one oligosaccharide.

[0219] In another and / or additional preferred embodiment, the oligosaccharides present in the sugar solution are selected from the list comprising: fucosylated oligosaccharides, neutral (uncharged) oligosaccharides, negatively charged oligosaccharides, sialylated oligosaccharides, Lewis-type antigens, oligosaccharides containing N-acetylglucosamine, neutral (uncharged) oligosaccharides containing N-acetylglucosamine, oligosaccharides containing N-acetyllactosamine, oligosaccharides containing lactose-N-disaccharide, oligosaccharides containing galactose, non-fucosylated neutral (uncharged) oligosaccharides, chitosan, oligosaccharides containing chitosan, heparin precursors, chondroitin sulfate, glycosaminoglycan oligosaccharides , heparin, heparan sulfate, dermatan sulfate, hyaluronan or hyaluronic acid, keratan sulfate, milk oligosaccharides, O-antigen, enterobacterial common antigen (ECA), oligosaccharide repeating units present in capsular polysaccharides, peptidoglycan, amino sugars, antigens of the human ABO blood group system, animal oligosaccharides, plant oligosaccharides, glucopyranosylsucrose (Glc-a1,4-Glc-a1,2-Fru), lactulose-N-trisaccharide II (GlcNAc-b1,3-Gal-b1,4-Fru), lactulose-N-tetraose, lactulose-N-neotetraose and uridine diphosphate galactose. In a more preferred embodiment, the fucosylated oligosaccharide is selected from the list comprising 2'-fucosyllactose (2'FL), 3-fucosyllactose (3FL), 4-fucosyllactose (4FL), 6-fucosyllactose (6FL), 2',3-difucosyllactose (diFL), lactose-N-fucopentaose I (LNFP I), Gal-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (Gal-LNFP I), GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (GalNAc-LNFP I), lactose-N-neofucopentaose I (LNnFP I), lactose-N-fucopentaose II (LNFP II), and lactose-N-fucopentaose II (LNFP II). II), lactose-N-fucopentaose III (LNFP III), lactose-N-fucopentaose V (LNFP V), lactose-N-fucopentaose VI (LNFP VI), lactose-N-neofucopentaose V, lactose-N-difucohexaose I (LNDFH-I), lactose-N-difucohexaose II (LNDFH-II), Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,4-(Fuc-a1,2-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, monofucosyllactose-N-hexaose-III, difucosyllactose-N-hexaose (a), difucosyllactose-N-hexaose, difucosyllactose-N-neohexaose, trifucosyllactose-N-hexaose, a 1,3-Galactosyl-3-fucosyllactose, Gal-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, 2-fucosyllactulose, 3-fucosyl-N-acetyllactosamine, 2'-fucosyl-N-acetyllactosamine, difucosyl-N-acetyllactosamine, 4-fucosyllactose-N-disaccharide, 2'-fucosyllactose-N-disaccharide, difucosyllactose-N-disaccharide, and GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc. In a more preferred embodiment, the sialylated oligosaccharide is selected from 3' sialyllactose (3'SL), 6' sialyllactose (6'SL), 8' sialyllactose (8'SL), 3,6-disialyllactose (Neu5Ac-α2,3-(Neu5Ac-α2,6)-Gal-β1,4-Glc), 6,6'-disialyllactose (Neu5Ac-α2,6-Gal-β1,4-(Neu5Ac-α2,6)-Glc), 8,3-disialyllactose (Neu5Ac-α2,8-Neu5Ac-α2,3-Gal-β1,4-Glc).4-Glc), 6'-sialyllactosamine, 3'-sialyllactosamine, sialyllactose-N-tetraose a (LSTa), sialyllactose-N-tetraose b (LSTb), sialyllactose-N-tetraose c (LSTc), sialyllactose-N-tetraose d (LSTd), disialyllactose-N-tetraose, disialyllactose-N-neotetraose, monosialyllactose-N-hexaose, disialyllactose-N-hexaose I, disialyllactose-N-hexaose II, monosialyllactose-N-neohexaose I, monosialyllactose-N-neohexaose II, disialyllactose-N-neohexaose, 3'-sialyl-3-fucosyllactose, fucodisialyllactose-N-hexaose, disialylmonofucosyllactose- N-neohexaose, sialyllactose-N-fucohexaose II, disialyllactose-N-fucopentaose II, monofucosyldisialyllactose-N-tetraose, Neu5Ac-a2,3-Gal-b1,4-GlcNAc-b1,3-Gal, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-KDO-lactose, 3'-KDO-lactosamine, 3'-KDO-6' sialyllactose, 3'KDO-8-sialyllactose, KDO-2,3Galβ-1,3GalNacβ-1,3Galα-1,4Galβ-1,4Gal, KDO-2,3Galβ-1,3GlcNacβ-1, 3Galβ-1,4Glc, KDO-2,3Galβ-1,4GlcNacβ-1,3Galβ-1,4Glc, 3'-KDO-3-fucosyllactose, Neu5Ac-a2,8-Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-sialyl-2'-fucosyllactose, 6'-sialyl-2'-fucosyllactose, 6'-sialyl-3-fucosyllactose, 3'-sialyl-3-fucosyllactose, Neu5Ac-a2,6-(Neu5Ac-a2,3-)Gal-b1,4-Glc, 3'-sialyl-3-fucosyllactosamine, Fuc-a 1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc, 6'-sialyllactose-N-disaccharide, 3'-sialyllactose-N-disaccharide, Neu5Ac-a2,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,3-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5 Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal -b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-( Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-)Gal- b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Fuc-a1,4-(Neu5Ac-a2,3-Gal -b1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fu c-a1,3-)Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, and any combination thereof. In another and / or additionally more preferred embodiment, the neutral (uncharged) oligosaccharide containing N-acetylglucosamine is selected from lacto-N-triose II (LN3), lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), 6'-galactosyl lactose, 4'-galactosyl lactose, 3'-galactosyl lactose, GlcNAc-b1,6-Gal-b1,4-Glc, lacto-N-hexaose (LNH), lacto-N-neohexaose (LNnH), p-lacto-N-hexaose (pLNH), p-lacto-N-neohexaose (pLNnH), GlcNAc-b1,6-(GlcNAc-b1,3-)Gal-b1,4-Glc4-Glc, lactose-N-pentaose (LN5), lactose-N-neopentaose, p-lactose-N-pentaose, p-lactose-N-neopentaose, lactose-N-novopentaose I, lactose-N-heptaose (LN7), lactose-N-neoheptaose, p-lactose-N-neoheptaose, p-lactose-N-heptaose, lactose-N-octaose (LNO), lactose-N-neooctaose, allolactose-N-octaose, p-lactose-N-octaose, allolactose-N-neooctaose, novel lactose-N-neooctaose, p-lactose-N-neooctaose (pLNnO), allolactose-N-nonaose, novel lactose-N-nonaose, lactose-N-nonaose (LN9) In another and / or additionally more preferred embodiment, the milk oligosaccharide is a mammalian milk oligosaccharide (MMO), more preferably a human milk oligosaccharide (HMO). In another and / or additionally more preferred embodiment, the animal oligosaccharide is selected from N-glycans and O-glycans. In another and / or additionally more preferred embodiment, the plant oligosaccharide is selected from N-glycans and O-glycans.

[0220] In another and / or additional preferred embodiment, the sugar composition further comprises one or more disaccharides.

[0221] In additional and / or alternative preferred embodiments, the sugar composition further comprises a protein source, vitamins, oils, minerals, fatty acids, enzymes and / or probiotic strains.

[0222] In another and / or additional preferred embodiment, the sugar composition is a composition selected from the group consisting of a food composition, a pharmaceutical composition, a cosmetic composition, a nutritional composition, a liquid food composition, a solid food composition, an infant food formula, a toddler food formula, a medical nutritional product, a medical food composition, a dietary supplement, a sachet product, a liquid ready-to-use infant nutritional product, a solid infant nutritional product, a liquid ready-to-use toddler nutritional product, a granulated product, a spray-dried infant formula, a premix, a milk drink, an infant formula, and combinations thereof.

[0223] Unless otherwise defined, all technical and scientific terms used herein generally have the same meaning as those commonly understood by those of ordinary skill in the art to which the invention pertains. Generally, the nomenclature used herein and the laboratory procedures in cell culture, molecular genetics, organic chemistry and nucleic acid chemistry, and hybridization described above and below are those well known and commonly used in the art. Standard techniques are used for nucleic acid and peptide synthesis. Generally, purification steps are performed according to the manufacturer's instructions.

[0224] Further advantages are apparent from the specific embodiments and examples. It goes without saying that the features mentioned above and those still to be explained below can be used not only in the respectively specified combination but also in other combinations or alone without departing from the scope of the present invention.

[0225] Furthermore, the present invention relates to the following specific embodiments:

[0226] 1. A method for purifying oligosaccharides from a solution, wherein the solution containing the oligosaccharides is a solution selected from the group consisting of a biocatalytic reaction solution, a chemical synthesis solution, and a cell culture, wherein the oligosaccharides are produced by the biocatalytic reaction solution, the chemical synthesis solution, or by cells cultured in the cell culture, the method comprising subjecting the solution to a reaction mixture containing H + The method comprises the step of: first cation exchange with a cation exchange resin in the form of a cation exchange resin, thereby converting the salt in the solution into an acid completely or partially, characterized in that the solution containing the salt converted into an acid completely or partially is passed through a cation exchange resin containing Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + The second cation exchange of the cation exchange resin in the form of Na + form.

[0227] 2. The method according to embodiment 1, wherein the cation exchange resin in the first and / or second cation exchange is selected from the list consisting of weak acid cation (WAC) exchange resins and strong acid cation (SAC) exchange resins.

[0228] 3. The method according to any one of embodiments 1 or 2, wherein the cation exchange resin in the first and / or second cation exchange has an acrylic-based, methacrylic-based, styrene-based or polystyrene-based matrix.

[0229] 4. The method according to embodiment 3, wherein the matrix further comprises divinylbenzene (DVB).

[0230] 5. The process according to any one of the preceding embodiments, wherein the flow rate through the first and / or second cation exchange is at least 0.5 bed volumes per hour (BV / h), preferably at least 1 BV / h, more preferably at least 2 BV / h, more preferably at least 2.5 BV / h, and most preferably at least 3 BV / h.

[0231] 6. The method according to any of the preceding embodiments, wherein the first and / or second cation exchange is performed at a temperature of 0°C to 80°C, preferably 4°C to 60°C, more preferably 4°C to 40°C, even more preferably 10°C to 37°C, even more preferably 20°C to 30°C, even more preferably 20°C to 25°C, even more preferably 22°C to 24°C, most preferably 23°C to 24°C.

[0232] 7. The method according to any one of the preceding embodiments, wherein the first and / or second cation exchange is performed at a pH in the range of 2 to 5, preferably 3 to 5, more preferably 4 to 5.

[0233] 8. The method according to any one of the preceding embodiments, wherein the method further comprises any one or more processes selected from the group consisting of homogenization, clarification, clearing, concentration, centrifugation, decantation, dilution, pH adjustment, temperature adjustment, filtration, ultrafiltration, microfiltration, diafiltration, reverse osmosis, electrodialysis, electrodeionization, nanofiltration, dialysis, use of activated carbon or carbon, use of a solvent, use of an alcohol, use of an aqueous alcohol mixture, use of charcoal, tangential flow high performance filtration, tangential flow ultrafiltration, affinity chromatography, ion exchange, ion exchange chromatography, mixed bed ion exchange, hydrophobic interaction chromatography, gel filtration, ligand exchange chromatography, column chromatography, cation exchange adsorption resins, anion exchange adsorption resins, use of an adsorbent material, use of an ion exchange resin, evaporation, wiped film evaporation, falling film evaporation, pasteurization, enzymatic treatment, decolorization, and drying, and wherein any one or more of the processes (a) is carried out in a reaction mixture comprising H + Before the first cation exchange of the cation exchange resin in the form of (a), and / or any one or more of the processes (b) comprising Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + form, preferably Na + After said second cation exchange of the cation exchange resin in the form of cation exchange resin, it is preferred that any one or more of said processes is performed more than once during said process.

[0234] 9. The method according to any one of the preceding embodiments, wherein the method further comprises any one or more processes selected from the group consisting of homogenization, clarification, clearing, concentration, centrifugation, decantation, dilution, pH adjustment in the range of 2 to 5, temperature adjustment, filtration, ultrafiltration, microfiltration, diafiltration, reverse osmosis, nanofiltration, dialysis, use of activated carbon or carbon, use of a solvent in the range of pH 2 to 5, use of an alcohol, use of an aqueous alcohol mixture, use of charcoal, tangential flow high performance filtration, tangential flow ultrafiltration, affinity chromatography, hydrophobic interaction chromatography, gel filtration, ligand exchange chromatography, column chromatography, cation exchange adsorption resins, use of an adsorbent material, evaporation, wiped film evaporation, falling film evaporation, pasteurization, enzymatic treatment, decolorization, drying, and dissolution in a liquid in the range of pH 2 to 5, and wherein any one or more of the processes is carried out in a liquid comprising H + After the first cation exchange of the cation exchange resin in the form of Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + form, preferably Na + Form a cation exchange resin before the second cation exchange.

[0235] 10. The method according to any one of the preceding embodiments, wherein the temperature of the solution is adjusted to a temperature of 36°C to 65°C, wherein the temperature is within 5°C of the temperature at which the solution exhibits maximum turbidity, preferably 36°C to 60°C, more preferably 40°C to 55°C, even more preferably 40°C to 45°C.

[0236] 11. The method according to any one of the preceding embodiments, wherein the temperature of the solution is adjusted to a temperature of 0°C to 122°C, preferably 2°C to 80°C, more preferably 4°C to 60°C, even more preferably 10°C to 55°C, even more preferably 20°C to 45°C, even more preferably 21°C to 40°C, even more preferably 22°C to 37°C, even more preferably 25°C to 30°C.

[0237] 12. The method according to any one of embodiments 8 to 11, wherein the method comprises two consecutive nanofiltration steps.

[0238] 13. The method according to any one of embodiments 8 to 12, wherein the method comprises two consecutive electrodialysis steps.

[0239] 14. The method of any one of embodiments 8 to 12, wherein the method does not comprise electrodialysis.

[0240] 15. The method according to any one of embodiments 8 to 14, wherein the method comprises two consecutive ultrafiltration steps, wherein the membrane molecular weight cut-off used in the first ultrafiltration step is higher than the membrane molecular weight cut-off used in the second ultrafiltration step.

[0241] 16. The method of any one of embodiments 8 to 15, wherein the method comprises clarification by any one or more of microfiltration, centrifugation, flocculation, or ultrafiltration.

[0242] 17. The method of any one of embodiments 8 to 16, wherein the method comprises drying selected from the list consisting of spray drying, freeze drying, spray freeze drying, freeze spray drying, belt drying, conveyor belt drying, vacuum belt drying, vacuum conveyor belt drying, drum drying, roller drying, vacuum drum drying, vacuum roller drying, and stirred film drying.

[0243] 18. The method according to any one of embodiments 8 to 17, wherein the method comprises filtration by using a filter aid and / or a flocculant, preferably the filter aid is an adsorbent, more preferably activated carbon.

[0244] 19. The method of any one of embodiments 8 to 18, wherein the method comprises ultrafiltration, wherein the ultrafiltration has a molecular weight cut-off equal to or higher than 1 kDa, 2 kDa, 3 kDa, 4 kDa, 5 kDa, 6 kDa, 7 kDa, 8 kDa, 9 kDa, 10 kDa, 11 kDa, 12 kDa, 13 kDa, 14 kDa, 15 kDa.

[0245] 20. The method according to any one of embodiments 8 to 19, wherein the method comprises nanofiltration, wherein the nanofiltration membrane used in the nanofiltration has a thickness of ≤ size exclusion limit.

[0246] 21. The method according to any one of embodiments 8 to 20, wherein the method comprises diafiltration of the solution until a conductivity of ≤ 15 mS / cm, preferably ≤ 10 mS / cm, ≤ 5 mS / cm, ≤ 1 mS / cm, ≤ 0.1 mS / cm, ≤ 0.01 mS / cm, ≤ 0.001 mS / cm is reached.

[0247] 22. The method of any one of embodiments 8 to 21, wherein the method comprises an enzymatic treatment comprising incubating the solution with one or more enzymes selected from the group consisting of glycosidase, lactase, β-galactosidase, fucosidase, sialidase, maltase, amylase, hexaminase, glucuronidase, trehalase, and invertase.

[0248] 23. The method of any one of embodiments 8 to 22, wherein the method comprises enzymatic treatment to convert lactose, sucrose, malto-oligosaccharides, maltotriose, sorbitol, trehalose, starch, cellulose, hemicellulose, lignocellulose, molasses, corn syrup and / or high fructose syrup into monosaccharides.

[0249] 24. The process according to any one of the preceding embodiments, wherein the process is a batch or continuous process.

[0250] 25. The method according to any one of the preceding embodiments, wherein the oligosaccharide is selected from the list comprising: fucosylated oligosaccharides, neutral (uncharged) oligosaccharides, negatively charged oligosaccharides, sialylated oligosaccharides, Lewis-type antigens, oligosaccharides containing N-acetylglucosamine, neutral (uncharged) oligosaccharides containing N-acetylglucosamine, oligosaccharides containing N-acetyllactosamine, oligosaccharides containing lactose-N-disaccharide, oligosaccharides containing galactose, non-fucosylated neutral (uncharged) oligosaccharides, chitosan, chitosan-containing oligosaccharides, heparin precursors, chondroitin sulfate, glycosaminoglycan oligosaccharides, heparin, heparan sulfate, chondroitin sulfate, dermatan sulfate, hyaluronan or hyaluronic acid, keratan sulfate, milk oligosaccharides, O-antigens, enterobacterial common antigens (ECA), Oligosaccharide repeating units present in capsular polysaccharides, peptidoglycans, amino sugars, antigens of the human ABO blood group system, animal oligosaccharides, plant oligosaccharides, glucopyranosylsucrose (Glc-a1, 4-Glc-a1, 2-Fru), lactulose-N-triose II (GlcNAc-b1, 3-Gal-b1, 4-Fru), lactulose-N-tetraose, lactulose-N-neotetraose and uridine diphosphate galactose; preferably wherein the fucosylated oligosaccharide is selected from the list comprising: 2'-fucosyllactose (2'FL), 3-fucosyllactose (3FL), 4-fucosyllactose (4FL), 6-fucosyllactose (6FL), 2', 3-difucosyllactose (diFL), lactose-N-fucopentaose I (LNFP I), Gal-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (Gal-LNFPI), GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (GalNAc-LNFP I), lactose-N-neofucopentaose I (LNnFP I), lactose-N-fucopentaose II (LNFP II), lactose-N-fucopentaose III (LNFPIII), lactose-N-fucopentaose V (LNFP V), lactose-N-fucopentaose VI (LNFP VI), lactose-N-neofucopentaose V, lactose-N-difucohexaose I (LNDFH-I), lactose-N-difucohexaose II (LNDFH-II), Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1 ,4-(Fuc-a1,3-)Glc, Fuc-a1,4-(Fuc-a1,2-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, monophylla Glycosyllactose-N-hexaose-III, difucosyllactose-N-hexaose (a), difucosyl-lactose-N-hexaose, difucosyl-lactose-N-neohexaose, trifucosyllactose-N-hexaose, a1,3-galactosyl-3-fucosyllactose, Gal-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a 1,3-)Glc, 2-fucosyllactulose, 3-fucosyl-N-acetyllactosamine, 2'-fucosyl-N-acetyllactosamine, difucosyl-N-acetyllactosamine, 4-fucosyllactose-N-disaccharide, 2'-fucosyllactose-N-disaccharide, difucosyllactose-N-disaccharide and GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc; preferably, wherein the sialylated oligosaccharide is selected from 3' sialyllactose (3'SL), 6'-sialyllactose (6'SL), 8'-sialyllactose (8'SL), 3,6-disialyllactose (Neu5Ac-α2,3-(Neu5Ac-α2,6)-Gal-β1,4-Glc), 6,6'-disialyllactose (Neu5Ac-α2,6-Gal-β1,4-(Neu5Ac-α2,6)-Glc), 8,3-disialyllactose (Neu5Ac-α2,8-Neu5Ac-α2,3-Gal-β1,4-Glc), 6'-sialyllactosamine, 3'-sialyllactosamine, sialyllactose-N-tetraose a (LSTa), sialyllactose-N-tetraose b (LSTb), sialyllactose-N-tetraose c (LSTc), sialyllactose-N-tetraose d (LSTd), disialyllactose-N-tetraose, disialyllactose-N-neotetraose, monosialyllactose-N-hexaose, disialyllactose-N-hexaose I, disialyllactose-N-hexaose II, monosialyllactose-N-neohexaose I, monosialyllactose-N-neohexaose II, disialyllactose-N-neohexaose, 3'-sialyl-3-fucosyllactose, fucodisialyllactose-N-hexaose, disialylmonofucosyllactose- N-neohexaose, sialyllactose-N-fucohexaose II, disialyllactose-N-fucopentaose II, monofucosyldisialyllactose-N-tetraose, Neu5Ac-a2,3-Gal-b1,4-GlcNAc-b1,3-Gal, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-KDO-lactose, 3'-KDO-lactosamine, 3'-KDO-6' sialyllactose, 3'KDO-8-sialyllactose, KDO-2,3Galβ-1,3GalNacβ-1,3Galα-1,4Galβ-1,4Gal, KDO-2,3Galβ-1,3GlcNacβ-1, 3Galβ-1,4Glc, KDO-2,3Galβ-1,4GlcNacβ-1,3Galβ-1,4Glc, 3'-KDO-3-fucosyllactose, Neu5Ac-a2,8-Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-sialyl-2'-fucosyllactose, 6'-sialyl-2'-fucosyllactose, 6'-sialyl-3-fucosyllactose, 3'-sialyl-3-fucosyllactose, Neu5Ac-a2,6-(Neu5Ac-a2,3-)Gal-b1,4-Glc, 3'-sialyl-3-fucosyllactosamine, Fuc-a 1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc, 6'-sialyllactose-N-disaccharide, 3'-sialyllactose-N-disaccharide, Neu5Ac-a2,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,3-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1, 3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Ga l-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-Gal-b1,3-GlcNAc-b1 ,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc-b1,3-G al-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-(Fuc-a1,2-) Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal -b1,4-Glc and any combination thereof; preferably wherein the neutral (uncharged) oligosaccharide containing N-acetylglucosamine is selected from lactose-N-triose II (LN3), lactose-N-tetraose (LNT), lactose-N-neotetraose (LNnT), 6'-galactosyl lactose, 4'-galactosyl lactose, 3'-galactosyl lactose, GlcNAc-b1,6-Gal-b1,4-Glc, lactose-N-hexaose (LNH), lactose-N-neohexaose (LNnH), p-lactose-N-hexaose (pLNH), p-lactose-N-neohexaose (pLNnH), GlcNAc-b1,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, lactose-N-hexaose (LNH), lactose-N-neohexaose (LNnH), p-lactose-N-hexaose (pLNH), p-lactose-N-neohexaose (pLNnH), GlcNAc-b1,6-(GlcNAc-b1,3-)Gal-b1,4-Glc4-Glc, lactose-N-pentaose (LN5), lactose-N-neopentaose, p-lactose-N-pentaose, p-lactose-N-neopentaose, lactose-N-novopentaose I, lactose-N-heptaose (LN7), lactose-N-neoheptaose, p-lactose-N-neoheptaose, p-lactose-N-heptaose, lactose-N-octaose (LNO), lactose-N-neooctaose, allolactose-N-octaose, p-lactose-N-octaose, allolactose-N-neooctaose, novel lactose-N-neooctaose, p-lactose-N-neooctaose (pLNnO), allolactose-N-nonaose, novel lactose-N-nonaose, lactose-N-nonaose (LN9), lactose-N-decaose, allolactose-N-decaose, novel lactose-N-decaose, lactose-N-neodecaose, p-lactose Glyco-N-neodecaose (pLNnD), α1,3-galactosyl lactose-N-neotetraose, GlcNAc-b1,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, GlcNAc-b1,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc and GlcNAc-b1,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc; preferably wherein the milk oligosaccharide is a mammalian milk oligosaccharide (MMO); more preferably human milk oligosaccharide (HMO); preferably wherein the animal oligosaccharide is selected from N-glycans and O-glycans; preferably wherein the plant oligosaccharide is selected from N-glycans and O-glycans.

[0251] 26. The method of any one of the preceding embodiments, wherein the oligosaccharide is a sialylated oligosaccharide having at least one sialic acid group selected from the group consisting of Neu4Ac; Neu5Ac; Neu4,5Ac2; Neu5,7Ac2; Neu5,8Ac2; Neu5,9Ac2; Neu4,5,9Ac3; Neu5,7,9Ac3; Neu5,8,9Ac3; Neu4,5,7,9Ac4; Neu5,7,8,9Ac4; Neu4,5,7,8,9Ac5; Neu5Gc and 2-keto-3-deoxymannose-octanone (KDO).

[0252] 27. The method of any one of the preceding embodiments, wherein the purity of the oligosaccharides in the solution, prior to purification by the method, is <70%, <60%, <50%, <40%, <30%, <20%, <10% based on total dry solids.

[0253] 28. The method according to any one of the preceding embodiments, wherein the oligosaccharide is accompanied in the solution by

[0254] - sialic acid,

[0255] - ash, preferably, said ash contains sulfate, phosphate, sodium, chloride, potassium, heavy metals, preferably said heavy metals contain ammonium, lead, arsenic, cadmium and / or mercury,

[0256] - one or more monosaccharides,

[0257] - one or more activated monosaccharides,

[0258] - one or more phosphorylated monosaccharides,

[0259] - one or more disaccharides, and / or

[0260] One or more oligosaccharides selected from the group consisting of neutral (uncharged) oligosaccharides, negatively charged oligosaccharides, milk oligosaccharides, preferably mammalian milk oligosaccharides (MMO), more preferably human milk oligosaccharides (HMO); O-antigens; enterobacterial common antigens (ECA); oligosaccharide repeating units present in capsular polysaccharides; peptidoglycans; amino sugars; Lewis-type antigenic oligosaccharides; antigens of the human ABO blood group system; animal oligosaccharides, preferably selected from N-glycans and O-glycans; plant oligosaccharides, preferably selected from plant oligosaccharides of N-glycans and O-glycans; glucopyranosylsucrose (Glc-a1 ,4-Glc-a1,2-Fru); lactulose-N-triose II (GlcNAc-b1,3-Gal-b1,4-Fru); lactulose-N-tetraose; lactulose-N-neotetraose and uridine diphosphate galactose; fucosylated oligosaccharides, which are preferably selected from the following list: 2'-fucosyllactose (2'FL), 3-fucosyllactose (3FL), 4-fucosyllactose (4FL), 6-fucosyllactose (6FL), 2',3-difucosyllactose (diFL), lactose-N-fucopentaose I (LNFP I), Gal-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (Gal-LNFP I), GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (GalNAc-LNFP I), lactose-N-neofucopentaose I (LNnFP I), lactose-N-fucopentaose II (LNFP II), lactose-N-fucopentaose III (LNFPIII), lactose-N-fucopentaose V (LNFP V), lactose-N-fucopentaose VI (LNFP VI), lactose-N-neofucopentaose V, lactose-N-difucohexaose I (LNDFH-I), lactose-N-difucohexaose II (LNDFH-II), Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1, 4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc,Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Gl c. Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,4-(Fuc-a1,2-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, monofucosyllactose-N-hexaose-III, difucosyllactose-N-hexaose (a), difucosyl-lactose-N-hexaose, difucosyl-lactose-N-neohexaose, trifucosyllactose-N-hexaose, a1,3-galactosyl-3-fucosyllactose, Gal-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, 2-fucosyllactulose, 3-fucosyl-N-acetyllactosamine , 2'-fucosyl-N-acetyllactosamine, difucosyl-N-acetyllactosamine, 4-fucosyllactose-N-disaccharide, 2'-fucosyllactose-N-disaccharide, difucosyllactose-N-disaccharide and GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc; sialylated oligosaccharides, which are preferably selected from 3' sialyllactose (3'SL), 6' sialyllactose (6'SL), 8' sialyllactose (8'SL), 3,6-disialyllactose (Neu5Ac-α2,3-(Neu5Ac-α2,6)-Gal-β1,4-Glc), 6,6'-disialyllactose (Neu5Ac-α2,6-Gal-β1,4-( Neu5Ac-α2,6)-Glc), 8,3-disialyl lactose (Neu5Ac-α2,8-Neu5Ac-α2,3-Gal-β1,4-Glc), 6'-sialyllactosamine, 3'-sialyllactosamine, sialyllactose-N-tetraose a (LSTa), sialyllactose-N-tetraose b (LSTb), sialyllactose-N-tetraose c (LSTc), sialyllactose-N-tetraose d (LSTd), disialyl lactose-N-tetraose, disialyl lactose-N-neotetraose, monosialyllactose-N-hexaose, disialyl lactose-N-hexaose I, disialyl lactose-N-hexaose II, monosialyllactose-N-neohexaose I, monosialyllactose-N-neohexaose I I, disialyl lactose-N-neohexaose, 3'-sialyl-3-fucosyllactose, fucodisialyl lactose-N-hexaose, disialyl monofucosyllactose-N-neohexaose, sialyl lactose-N-fucohexaose II, disialyl lactose-N-fucopentaose II, monofucosyl disialyl lactose-N-tetraose, Neu5Ac-a2,3-Gal-b1,4-GlcNAc-b1,3-Gal, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-KDO-lactose, 3'-KDO-lactosamine, 3'-KDO-6' sialyllactose, 3'KDO-8-sialyllactose, KDO-2,3Galβ-1,3GalNacβ-1,3Galα-1,4Galβ-1,4Gal, KDO-2,3Galβ-1,3GlcNacβ-1,3Galβ-1,4Glc, KDO-2,3Galβ-1,4GlcNacβ-1,3Galβ-1,4Glc, 3'-KDO-3-fucosyllactose, Neu5Ac-a2,8-Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-sialyl-2'-fucosyllactose, 6'-sialyl-2'-fucosyllactose, 6'-sialyl-3-fucosyllactose, 3'-sialyl-3-fucosyllactose, Neu5A c-a2,6-(Neu5Ac-a2,3-)Gal-b1,4-Glc, 3'-sialyl-3-fucosyllactosamine, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc, 6'-sialyl lactose-N-disaccharide, 3'-sialyl lactose-N-disaccharide, Neu5Ac-a2,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,3-Gal-b1,4-(Fuc-a1,3- )GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-GlcNA c-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(F uc-a1,3-)Glc, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-G lc, Neu5Ac-a2,6-(Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3 -)Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal- b1,4-Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc and any combination thereof; N-acetylglucosamine-containing oligosaccharides, which are preferably selected from lactose-N-triose II (LN3), lactose-N-tetraose (LNT), lactose-N-neotetraose (LNnT), 6'-galactosyl lactose, 4'-galactosyl lactose, 3'-galactosyl lactose, GlcNAc-b1,6-Gal-b1,4-Glc, lactose-N-hexaose (LNH), lactose-N-neohexose (LNnH), p-lactose-N-hexaose (pLNH), p-lactose-N-neohexose (pLNnH), GlcNAc-b1,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, lactose-N-pentaose (LN5), lactose-N-neopentaose, p-lactose-N-pentaose, p-lactose-N-neopentaose, lactose-N-novopentaose I, lactose-N-heptaose (LN7), lactose-N-neoheptaose, p-lactose-N-neoheptaose, p-lactose-N-heptaose, lactose-N-octaose (LNO), lactose-N-neooctaose, allolactose-N-octaose, p-lactose-N-octaose, allolactose-N-neooctaose, novel lactose-N-neooctaose, p-lactose-N-neooctaose (pLNnO), allolactose-N-nonaose , novel lactose-N-nonasaccharide, lactose-N-nonasaccharide (LN9), lactose-N-decaose, allolactose-N-decaose, novel lactose-N-decaose, lactose-N-neodecaose, para-lactose-N-neodecaose (pLNnD), a1,3-galactosyllactose-N-neotetraose, GlcNAc-b1,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, GlcNAc-b1,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc and GlcNAc-b1,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc; neutral (uncharged) oligosaccharides containing N-acetylglucosamine; oligosaccharides containing N-acetyllactosamine; oligosaccharides containing lactose-N-disaccharide; non-fucosylated neutral (uncharged) oligosaccharides; chitosan; chitosan-containing oligosaccharides; heparin precursors; chondroitin sulfate; glycosaminoglycan oligosaccharides; heparin; heparan sulfate; chondroitin sulfate; dermatan sulfate; hyaluronan; hyaluronic acid; and keratan sulfate.

[0261] 29. The method according to any one of the preceding embodiments, wherein the solution comprising the oligosaccharides prior to purification by the method has an ash content > 10% based on total dry solids.

[0262] 30. The method according to any one of the preceding embodiments, wherein the solution comprising the oligosaccharides prior to purification by the method has an ash content of >10% based on total dry solids, wherein the ash comprises any one or more heavy metals selected from the group consisting of lead, arsenic, cadmium, mercury, zinc, manganese, copper, iron, magnesium and calcium.

[0263] 31. The method according to any one of the preceding embodiments, wherein the solution comprising the oligosaccharides prior to purification by the method comprises a lead content of >0.1 mg / kg dry solids, an arsenic content of >0.2 mg / kg dry solids, a cadmium content of >0.1 mg / kg dry solids and / or a mercury content of >0.5 mg / kg dry solids.

[0264] 32. The method of any one of the preceding embodiments, wherein the solution is a cell culture using cells, preferably metabolically engineered cells, wherein the oligosaccharides are produced by the cells, the cell culture comprising the oligosaccharides, biomass, culture medium components and contaminants, wherein the purity of the oligosaccharides in the cell culture prior to purification by the method is <70%, <60%, <50%, <40%, <30%, <20%, <10% on a total dry solids basis.

[0265] 33. The method of any one of the preceding embodiments, wherein the solution is a cell culture using cells, preferably metabolically engineered cells, wherein the oligosaccharides are produced by the cells, the cell culture comprising the oligosaccharides, biomass, culture medium components and contaminants, wherein when present, biomass separated during the method is optionally recycled to the cell culture.

[0266] 34. The method according to any one of the preceding embodiments, wherein the solution is a cell culture using at least one cell that has been metabolically engineered to produce the oligosaccharide.

[0267] 35. A method according to any of the preceding embodiments, wherein the solution is a cell culture using at least one cell that has been metabolically engineered to produce the oligosaccharide and one or more of the following: i) sialic acid, ii) one or more monosaccharides, iii) one or more activated monosaccharides, iv) one or more phosphorylated monosaccharides, v) one or more disaccharides and / or vi) one or more other oligosaccharides.

[0268] 36. The method of any one of the preceding embodiments, wherein the cell produces the oligosaccharide from one or more internalized precursors.

[0269] 37. The method of any one of the preceding embodiments, wherein the cell is a bacterium, a fungus, a yeast, a plant cell, an animal cell, or a protozoan cell.

[0270] 38. The method of any one of the preceding embodiments, wherein the cells are cultured in a culture medium comprising a carbon source comprising monosaccharides, disaccharides, oligosaccharides, polysaccharides, polyols, glycerol, a complex culture medium comprising molasses, corn steep liquor, peptone, tryptone or yeast extract; preferably, the carbon source is selected from the group consisting of glucose, N-acetylglucosamine (GlcNAc), glycerol, fructose, sucrose, maltose, lactose, arabinose, maltooligosaccharides, maltotriose, sorbitol, xylose, rhamnose, galactose, mannose, methanol, ethanol, trehalose, starch, cellulose, hemicellulose, molasses, corn steep liquor, high fructose syrup, acetate, citrate, lactate and pyruvate; preferably, the culture medium is a chemically defined culture medium.

[0271] 39. The method of embodiment 38, wherein the culture medium is a minimal salt medium comprising sulfate, phosphate, chloride, ammonium, calcium, magnesium, sodium, potassium, iron, copper, zinc, manganese, cobalt and / or selenium.

[0272] 40. The method according to any one of the preceding embodiments, wherein the cell culture is fermentation.

[0273] 41. The method according to any one of the preceding embodiments, wherein the oligosaccharide is accompanied in the solution by one or more other oligosaccharides, wherein at least one of the other oligosaccharides has the same degree of polymerization (DP) as the oligosaccharide, preferably wherein all of the other oligosaccharides have the same DP as the oligosaccharide.

[0274] 42. The method according to any one of embodiments 1 to 40, wherein the oligosaccharide is accompanied in the solution by one or more other oligosaccharides, wherein at least one of the other oligosaccharides has a different degree of polymerization (DP) than the oligosaccharide, preferably wherein all of the other oligosaccharides have a different DP than the oligosaccharide.

[0275] 43. The method of any one of embodiments 28 to 42, wherein the oligosaccharides are purified from the sialic acid and / or the ash by said method.

[0276] 44. The method of any one of embodiments 28 to 42, wherein the oligosaccharides and the one or more other oligosaccharides are purified from the sialic acid and / or the ash by the method.

[0277] 45. The process according to any of the preceding embodiments, wherein the purity of the oligosaccharides obtained in the purified oligosaccharide solution at the end of the process is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, based on total dry solids.

[0278] 46. ​​The method according to any of the preceding embodiments, wherein the purified yield of oligosaccharides obtained in the purified oligosaccharide solution at the end of the method is at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%.

[0279] 47. The process according to any of the preceding embodiments, wherein the purified oligosaccharide solution obtained at the end of the process has an ash content of ≤10% based on total dry solids, preferably ≤9% based on total dry solids, more preferably ≤8% based on total dry solids, even more preferably ≤7% based on total dry solids, even more preferably ≤6% based on total dry solids, even more preferably ≤5% based on total dry solids, even more preferably ≤4% based on total dry solids, even more preferably ≤3% based on total dry solids, even more preferably ≤2% based on total dry solids, even more preferably ≤1% based on total dry solids, and most preferably ≤0.5% based on total dry solids.

[0280] 48. The process according to any one of the preceding embodiments, wherein the purified oligosaccharide solution obtained at the end of the process has an ash content, based on total dry solids, of ≤ 10%, wherein the ash comprises any one or more heavy metals selected from the group consisting of lead, arsenic, cadmium, mercury, zinc, manganese, copper, iron, magnesium and calcium.

[0281] 49. The process according to any of the preceding embodiments, wherein the purified oligosaccharide solution obtained at the end of the process has an ash content, based on total dry solids, of ≤ 10% and preferably has a lead content of less than 0.1 mg / kg dry solids, more preferably a lead content of less than 0.02 mg / kg dry solids, even more preferably a lead content of less than 0.01 mg / kg dry solids, an arsenic content of less than 0.2 mg / kg dry solids, more preferably an arsenic content of less than 0.05 mg / kg dry solids, even more preferably an arsenic content of less than 0.02 mg / kg dry solids, a cadmium content of less than 0.1 mg / kg dry solids, more preferably a cadmium content of less than 0.01 mg / kg dry solids and / or a mercury content of less than 0.5 mg / kg dry solids, more preferably a mercury content of less than 0.1 mg / kg dry solids, even more preferably a mercury content of less than 0.005 mg / kg dry solids.

[0282] 50. The process according to any of the preceding embodiments, wherein the purified oligosaccharide solution obtained at the end of the process is sterilized by filtration and / or subjected to endotoxin removal, preferably by filtration through a 3 kDa filter.

[0283] 51. The method according to any of the preceding embodiments, wherein the purified oligosaccharide solution obtained at the end of the method has a protein content of less than 100 mg / kg dry solids, a DNA content of less than 10 ng / g dry solids and / or an endotoxin content of less than 10,000 EU / g dry solids, preferably, the purified oligosaccharide solution is free of DNA, proteins and / or recombinant genetic material.

[0284] 52. The process according to any of the preceding embodiments, wherein the purified oligosaccharide solution obtained at the end of the process is further i) concentrated to a syrup of at least 20% dry matter, preferably at least 30% dry matter, more preferably at least 40% dry matter; ii) crystallized; iii) dried to a powder or iv) granulated.

[0285] 53. The process according to any of the preceding embodiments, wherein the purified oligosaccharide solution obtained at the end of the process is further concentrated to an oligosaccharide concentration of >100 g / L, preferably >200 g / L, more preferably >300 g / L, more preferably >400 g / L, more preferably >500 g / L, more preferably >600 g / L, most preferably from 300 g / L to 650 g / L and / or at a temperature of <80°C, preferably <60°C, more preferably <50°C, more preferably from 20°C to 50°C, even more preferably from 30°C to 45°C, preferably by a process comprising the use of vacuum evaporation or reverse osmosis or nanofiltration.

[0286] 54. The process according to any one of the preceding embodiments, wherein the purified oligosaccharide solution obtained at the end of the process comprises oligosaccharides concentrated to a concentration > 1.5 M and cooled to a temperature < 25°C, more preferably < 8°C, to obtain crystalline material of oligosaccharides.

[0287] 55. The method according to any one of embodiments 1 to 53, wherein the purified oligosaccharide solution obtained at the end of the method is dried by one or more drying steps selected from the group consisting of spray drying, freeze drying, evaporation, precipitation, spray freeze drying, freeze spray drying, belt drying, conveyor belt drying, vacuum belt drying, vacuum conveyor belt drying, drum drying, roller drying, vacuum drum drying, vacuum roller drying and stirred film drying.

[0288] 56. The method according to embodiment 55, wherein the purified oligosaccharide solution obtained at the end of the method is dried by spray drying, freeze drying or stirred thin film drying, and preferably wherein the pH of the purified oligosaccharide solution is from 2 to 7, preferably from 3 to 6, more preferably from 4 to 5.

[0289] 57. The process according to any one of embodiments 55 or 56, wherein the purified oligosaccharide solution obtained at the end of the process is dried by spray drying, preferably spray drying at an oligosaccharide solution concentration of 20-60 (w / v), preferably 30-50 (w / v), more preferably 35-45 (w / v), with a nozzle temperature of 110-150°C, preferably 120-140°C, more preferably 125-135°C and / or an outlet temperature of 60-80°C, preferably 65-70°C.

[0290] 58. A purified oligosaccharide solution, purified oligosaccharide or purified oligosaccharide mixture obtainable, preferably obtained, by a process according to any one of the preceding embodiments.

[0291] 59. Purified oligosaccharides obtainable, preferably obtained, by the process according to any one of embodiments 1 to 57, wherein the purified oligosaccharide solution comprising said purified oligosaccharides is i) dried, preferably spray dried or dried by means of an agitated thin film dryer; ii) freeze dried, iii) crystallized or iv) concentrated to a syrup of at least 20% dry matter, preferably at least 30% dry matter, more preferably at least 40% dry matter.

[0292] 60. A purified oligosaccharide mixture obtainable, preferably obtained, by a process according to any one of embodiments 1 to 57, wherein a purified oligosaccharide solution comprising said purified oligosaccharide mixture is i) dried, preferably spray dried or dried by means of an agitated thin film dryer; ii) freeze dried, iii) crystallized or iv) concentrated to a syrup of at least 20% dry matter, preferably at least 30% dry matter, more preferably at least 40% dry matter.

[0293] 61. Purifying oligosaccharides according to the method of any one of embodiments 1 to 57, wherein the purified oligosaccharides obtained after the method have an ash content of ≤ 10% based on total dry solids, preferably wherein the oligosaccharides are produced by cell culture.

[0294] 62. Oligosaccharides purified according to the method of any one of embodiments 1 to 57, and wherein the purified oligosaccharides obtained after the method have an ash content of ≤ 10% based on total dry solids, wherein the ash contains any one or more heavy metals selected from the group consisting of lead, arsenic, cadmium, mercury, zinc, manganese, copper, iron, magnesium and calcium.

[0295] 63. Oligosaccharides purified by the method according to any one of embodiments 1 to 57, wherein the purified oligosaccharides obtained after the method have a lead content of <0.1 mg / kg dry solid, preferably <0.02 mg / kg dry solid, more preferably <0.01 mg / kg dry solid; an arsenic content of <0.2 mg / kg dry solid, preferably <0.05 mg / kg dry solid, more preferably <0.02 mg / kg dry solid; a cadmium content of <0.1 mg / kg dry solid, preferably <0.01 mg / kg dry solid and / or a mercury content of <0.5 mg / kg dry solid, preferably <0.1 mg / kg dry solid, more preferably <0.005 mg / kg dry solid.

[0296] 64. A spray-dried oligosaccharide or oligosaccharide mixture, wherein the oligosaccharide or oligosaccharide mixture is purified according to the method of any one of embodiments 1 to 57, and wherein the spray-dried oligosaccharide or oligosaccharide mixture obtained after the method has an ash content of ≤10% based on total dry solids, preferably wherein the oligosaccharide or oligosaccharide mixture is produced by cell culture.

[0297] 65. A dry powder of the purified oligosaccharide solution obtained from the method of any one of embodiments 55 to 57, wherein the dry powder:

[0298] - contains ≤ 15 wt. % water, preferably ≤ 10 wt. % water, more preferably ≤ 7 wt. % water, most preferably ≤ 5 wt. % water, and / or

[0299] - having an average particle size of 50 to 250 μm, preferably 95 to 120 μm, more preferably 110 to 120 μm, wherein the particle size is determined by laser diffraction, preferably the powder is a spray-dried powder.

[0300] 66. The dry powder according to any one of embodiments 59, 60, 64, 65, wherein the powder, when redissolved in water at a concentration of 10% (mass / volume), provides a solution having a pH of 4 to 7, preferably a pH of 4 to 6, more preferably a pH of 4 to 5, even more preferably a pH of 5 to 6.

[0301] 67. The purified oligosaccharide according to any one of embodiments 58, 59, 61 to 65, wherein any one or more of the oligosaccharides are milk oligosaccharides, preferably mammalian milk oligosaccharides (MMOs), more preferably human milk oligosaccharides (HMOs).

[0302] 68. The purified oligosaccharide mixture comprising oligosaccharides according to any one of embodiments 58, 60 to 65, wherein the purified oligosaccharide mixture comprises milk oligosaccharides, preferably mammalian milk oligosaccharides (MMO), more preferably human milk oligosaccharides (HMO).

[0303] 69. The purified oligosaccharide or purified oligosaccharide mixture according to any one of embodiments 58 to 68, wherein the purified oligosaccharide or purified oligosaccharide mixture a) has a conductivity of less than 10 mS / cm at 300 g / L solution; b) is free of recombinant DNA material, optionally free of any DNA; and / or c) is free of proteins derived from a recombinant microorganism, optionally free of any protein.

[0304] 70. The purified oligosaccharide or purified oligosaccharide mixture according to any one of embodiments 58 to 69 for use in medicine, preferably for the prevention or treatment of gastrointestinal diseases.

[0305] 71. Use of a purified oligosaccharide obtained by the method of any one of embodiments 1 to 57 in a food or feed formulation, a dietary supplement, a cosmetic ingredient or a pharmaceutical ingredient.

[0306] 72. Use of the purified oligosaccharide according to any one of embodiments 58 to 69 in a food or feed formulation, a dietary supplement, a cosmetic ingredient or a pharmaceutical ingredient.

[0307] 73. The use according to any one of embodiments 71 or 72, wherein:

[0308] - the food is a human food, preferably a baby food, a human infant food and / or an infant formula or an infant supplement, and / or

[0309] - The feed is a pet food, an animal milk replacer, a veterinary product, a veterinary feed supplement, a nutritional supplement, a weaning feed or a creep feed.

[0310] 74. Use of the milk oligosaccharide according to any one of embodiments 67 or 68 as an additive in food, preferably as an additive in human food and / or pet food, more preferably as an additive in human infant food.

[0311] 75. A purified oligosaccharide comprising an ash content of ≤ 10% based on total dry solids, wherein the ash comprises any one or more heavy metals selected from the group consisting of lead, arsenic, cadmium, mercury, zinc, manganese, copper, iron, magnesium and calcium.

[0312] 76. The purified oligosaccharide according to embodiment 75, wherein the ash content of ≤ 10% based on total dry solids comprises a lead content of less than 0.1 mg / kg dry solids, preferably a lead content of less than 0.02 mg / kg dry solids, more preferably a lead content of less than 0.01 mg / kg dry solids, an arsenic content of less than 0.2 mg / kg dry solids, preferably an arsenic content of less than 0.05 mg / kg dry solids, more preferably an arsenic content of less than 0.02 mg / kg dry solids, a cadmium content of less than 0.1 mg / kg dry solids, preferably a cadmium content of less than 0.01 mg / kg dry solids and / or a mercury content of less than 0.5 mg / kg dry solids, preferably a mercury content of less than 0.1 mg / kg dry solids, more preferably a mercury content of less than 0.005 mg / kg dry solids.

[0313] 77. The purified oligosaccharide according to any one of embodiments 75 or 76, wherein the purified oligosaccharide is selected from the list comprising: fucosylated oligosaccharides, neutral (uncharged) oligosaccharides, negatively charged oligosaccharides, sialylated oligosaccharides, Lewis-type antigens, oligosaccharides containing N-acetylglucosamine, neutral (uncharged) oligosaccharides containing N-acetylglucosamine, oligosaccharides containing N-acetyllactosamine, oligosaccharides containing lactose-N-disaccharide, oligosaccharides containing galactose, non-fucosylated neutral (uncharged) oligosaccharides, chitosan, oligosaccharides containing chitosan, heparin precursors, chondroitin sulfate, glycosaminoglycan oligosaccharides, heparin, heparan sulfate, chondroitin sulfate, dermatan sulfate, hyaluronan or hyaluronic acid, keratan sulfate, milk oligosaccharides, O-antigens, enterobacterial common antigen (EC A), oligosaccharide repeating units present in capsular polysaccharides, peptidoglycans, amino sugars, antigens of the human ABO blood group system, animal oligosaccharides, plant oligosaccharides, glucopyranosylsucrose (Glc-a1,4-Glc-a1,2-Fru), lactulose-N-triose II (GlcNAc-b1,3-Gal-b1,4-Fru), lactulose-N-tetraose, lactulose-N-neotetraose and uridine diphosphate galactose; preferably wherein the fucosylated oligosaccharide is selected from the list comprising the following: 2'-fucosyllactose (2'FL), 3-fucosyllactose (3FL), 4-fucosyllactose (4FL), 6-fucosyllactose (6FL), 2',3-difucosyllactose (diFL), lactose-N-fucopentaose I (LNFP I), Gal-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (Gal-LNFP I), GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (GalNAc-LNFP I), lactose-N-neofucopentaose I (LNnFP I), lactose-N-fucopentaose II (LNFP II), lactose-N-fucopentaose III (LNFP III), lactose-N-fucopentaose V (LNFP V), lactose-N-fucopentaose VI (LNFP VI), lactose-N-neofucopentaose V, lactose-N-difucohexaose I (LNDFH-I), lactose-N-difucohexaose II (LNDFH-II), Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1 ,4-(Fuc-a1,3-)Glc, Fuc-a1,4-(Fuc-a1,2-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, monophylla Glycosyllactose-N-hexaose-III, difucosyllactose-N-hexaose (a), difucosyl-lactose-N-hexaose, difucosyl-lactose-N-neohexaose, trifucosyllactose-N-hexaose, a1,3-galactosyl-3-fucosyllactose, Gal-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a 1,3-)Glc, 2-fucosyllactulose, 3-fucosyl-N-acetyllactosamine, 2'-fucosyl-N-acetyllactosamine, difucosyl-N-acetyllactosamine, 4-fucosyllactose-N-disaccharide, 2'-fucosyllactose-N-disaccharide, difucosyllactose-N-disaccharide and GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc; preferably, wherein the sialylated oligosaccharide is selected from 3' sialyllactose (3'SL), 6'-sialyllactose (6'SL), 8'-sialyllactose (8'SL), 3,6-disialyllactose (Neu5Ac-α2,3-(Neu5Ac-α2,6)-Gal-β1,4-Glc), 6,6'-disialyllactose (Neu5Ac-α2,6-Gal-β1,4-(Neu5Ac-α2,6)-Glc), 8,3-disialyllactose (Neu5Ac-α2,8-Neu5Ac-α2,3-Gal-β1,4-Glc), 6'-sialyllactosamine, 3'-sialyllactosamine, sialyllactose-N-tetraose a (LSTa), sialyllactose-N-tetraose b (LSTb), sialyllactose-N-tetraose c (LSTc), sialyllactose-N-tetraose d (LSTd), disialyllactose-N-tetraose, disialyllactose-N-neotetraose, monosialyllactose-N-hexaose, disialyllactose-N-hexaose I, disialyllactose-N-hexaose II, monosialyllactose-N-neohexaose I, monosialyllactose-N-neohexaose II, disialyllactose-N-neohexaose, 3'-sialyl-3-fucosyllactose, fucodisialyllactose-N-hexaose, disialylmonofucosyllactose- N-neohexaose, sialyllactose-N-fucohexaose II, disialyllactose-N-fucopentaose II, monofucosyldisialyllactose-N-tetraose, Neu5Ac-a2,3-Gal-b1,4-GlcNAc-b1,3-Gal, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-KDO-lactose, 3'-KDO-lactosamine, 3'-KDO-6' sialyllactose, 3'KDO-8-sialyllactose, KDO-2,3Galβ-1,3GalNacβ-1,3Galα-1,4Galβ-1,4Gal, KDO-2,3Galβ-1,3GlcNacβ-1, 3Galβ-1,4Glc, KDO-2,3Galβ-1,4GlcNacβ-1,3Galβ-1,4Glc, 3'-KDO-3-fucosyllactose, Neu5Ac-a2,8-Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-sialyl-2'-fucosyllactose, 6'-sialyl-2'-fucosyllactose, 6'-sialyl-3-fucosyllactose, 3'-sialyl-3-fucosyllactose, Neu5Ac-a2,6-(Neu5Ac-a2,3-)Gal-b1,4-Glc, 3'-sialyl-3-fucosyllactosamine, Fuc-a 1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc, 6'-sialyllactose-N-disaccharide, 3'-sialyllactose-N-disaccharide, Neu5Ac-a2,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,3-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1, 3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Ga l-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-Gal-b1,3-GlcNAc-b1 ,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc-b1,3-G al-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-(Fuc-a1,2-) Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal -b1,4-Glc and any combination thereof; preferably wherein the neutral (uncharged) oligosaccharide containing N-acetylglucosamine is selected from lactose-N-triose II (LN3), lactose-N-tetraose (LNT), lactose-N-neotetraose (LNnT), 6'-galactosyl lactose, 4'-galactosyl lactose, 3'-galactosyl lactose, GlcNAc-b1,6-Gal-b1,4-Glc, lactose-N-hexaose (LNH), lactose-N-neohexaose (LNnH), p-lactose-N-hexaose (pLNH), p-lactose-N-neohexaose (pLNnH), GlcNAc-b1,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, lactose-N-hexaose (LNH), lactose-N-neohexaose (LNnH), p-lactose-N-hexaose (pLNH), p-lactose-N-neohexaose (pLNnH), GlcNAc-b1,6-(GlcNAc-b1,3-)Gal-b1,4-Glc4-Glc, lactose-N-pentaose (LN5), lactose-N-neopentaose, p-lactose-N-pentaose, p-lactose-N-neopentaose, lactose-N-novopentaose I, lactose-N-heptaose (LN7), lactose-N-neoheptaose, p-lactose-N-neoheptaose, p-lactose-N-heptaose, lactose-N-octaose (LNO), lactose-N-neooctaose, allolactose-N-octaose, p-lactose-N-octaose, allolactose-N-neooctaose, novel lactose-N-neooctaose, p-lactose-N-neooctaose (pLNnO), allolactose-N-nonaose, novel lactose-N-nonaose, lactose-N-nonaose (LN9), lactose-N-decaose, allolactose-N-decaose, novel lactose-N-decaose, lactose-N-neodecaose, p-lactose Glyco-N-neodecaose (pLNnD), α1,3-galactosyl lactose-N-neotetraose, GlcNAc-b1,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, GlcNAc-b1,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc and GlcNAc-b1,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc; preferably wherein the milk oligosaccharide is a mammalian milk oligosaccharide (MMO), more preferably a human milk oligosaccharide (HMO); preferably wherein the animal oligosaccharide is selected from N-glycans and O-glycans; preferably wherein the plant oligosaccharide is selected from N-glycans and O-glycans.

[0314] 78. A sugar composition comprising an ash content of ≤10% based on total dry solids and an oligosaccharide content of ≥90% based on total dry solids, wherein the ash contains any one or more heavy metals selected from the group consisting of lead, arsenic, cadmium, mercury, zinc, manganese, copper, iron, magnesium and calcium.

[0315] 79. The composition according to embodiment 78, wherein the ash content of ≤ 10% based on total dry solids comprises a lead content of less than 0.1 mg / kg dry solids, preferably a lead content of less than 0.02 mg / kg dry solids, more preferably a lead content of less than 0.01 mg / kg dry solids, an arsenic content of less than 0.2 mg / kg dry solids, preferably an arsenic content of less than 0.05 mg / kg dry solids, more preferably an arsenic content of less than 0.02 mg / kg dry solids, a cadmium content of less than 0.1 mg / kg dry solids, preferably a cadmium content of less than 0.01 mg / kg dry solids and / or a mercury content of less than 0.5 mg / kg dry solids, preferably a mercury content of less than 0.1 mg / kg dry solids, more preferably a mercury content of less than 0.005 mg / kg dry solids.

[0316] 80. The composition of any one of embodiments 78 or 79, wherein the composition comprises more than one oligosaccharide.

[0317] 81. The composition of any one of embodiments 78 to 80, wherein the oligosaccharide is selected from the list comprising: fucosylated oligosaccharides, neutral (uncharged) oligosaccharides, negatively charged oligosaccharides, sialylated oligosaccharides, Lewis-type antigens, N-acetylglucosamine-containing oligosaccharides, N-acetylglucosamine-containing neutral (uncharged) oligosaccharides, N-acetyllactosamine-containing oligosaccharides, lactose-N-disaccharide-containing oligosaccharides, galactose-containing oligosaccharides, non-fucosylated neutral (uncharged) oligosaccharides, chitosan, chitosan-containing oligosaccharides, heparin precursors, chondroitin sulfate, glycosaminoglycan oligosaccharides, heparin, heparan sulfate, chondroitin sulfate, dermatan sulfate, hyaluronan or hyaluronic acid, keratan sulfate, milk oligosaccharides, O-antigens, enterobacterial common antigens (ECAs), ), oligosaccharide repeating units present in capsular polysaccharides, peptidoglycans, amino sugars, antigens of the human ABO blood group system, animal oligosaccharides, plant oligosaccharides, glucopyranosylsucrose (Glc-a1,4-Glc-a1,2-Fru), lactulose-N-triose II (GlcNAc-b1,3-Gal-b1,4-Fru), lactulose-N-tetraose, lactulose-N-neotetraose and uridine diphosphate galactose; preferably wherein the fucosylated oligosaccharide is selected from the list comprising: 2'-fucosyllactose (2'FL), 3-fucosyllactose (3FL), 4-fucosyllactose (4FL), 6-fucosyllactose (6FL), 2',3-difucosyllactose (diFL), lactose-N-fucopentaose I (LNFP I), Gal-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (Gal-LNFP I), GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (GalNAc-LNFP I), lactose-N-neofucopentaose I (LNnFP I), lactose-N-fucopentaose II (LNFP II), lactose-N-fucopentaose III (LNFP III), lactose-N-fucopentaose V (LNFP V), lactose-N-fucopentaose VI (LNFP VI), lactose-N-neofucopentaose V, lactose-N-difucohexaose I (LNDFH-I), lactose-N-difucohexaose II (LNDFH-II), Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1 ,4-(Fuc-a1,3-)Glc, Fuc-a1,4-(Fuc-a1,2-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, monophylla Glycosyllactose-N-hexaose-III, difucosyllactose-N-hexaose (a), difucosyl-lactose-N-hexaose, difucosyl-lactose-N-neohexaose, trifucosyllactose-N-hexaose, a1,3-galactosyl-3-fucosyllactose, Gal-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a 1,3-)Glc, 2-fucosyllactulose, 3-fucosyl-N-acetyllactosamine, 2'-fucosyl-N-acetyllactosamine, difucosyl-N-acetyllactosamine, 4-fucosyllactose-N-disaccharide, 2'-fucosyllactose-N-disaccharide, difucosyllactose-N-disaccharide and GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc; preferably, wherein the sialylated oligosaccharide is selected from 3' sialyllactose (3'SL), 6'-sialyllactose (6'SL), 8'-sialyllactose (8'SL), 3,6-disialyllactose (Neu5Ac-α2,3-(Neu5Ac-α2,6)-Gal-β1,4-Glc), 6,6'-disialyllactose (Neu5Ac-α2,6-Gal-β1,4-(Neu5Ac-α2,6)-Glc), 8,3-disialyllactose (Neu5Ac-α2,8-Neu5Ac-α2,3-Gal-β1,4-Glc), 6'-sialyllactosamine, 3'-sialyllactosamine, sialyllactose-N-tetraose a (LSTa), sialyllactose-N-tetraose b (LSTb), sialyllactose-N-tetraose c (LSTc), sialyllactose-N-tetraose d (LSTd), disialyllactose-N-tetraose, disialyllactose-N-neotetraose, monosialyllactose-N-hexaose, disialyllactose-N-hexaose I, disialyllactose-N-hexaose II, monosialyllactose-N-neohexaose I, monosialyllactose-N-neohexaose II, disialyllactose-N-neohexaose, 3'-sialyl-3-fucosyllactose, fucodisialyllactose-N-hexaose, disialylmonofucosyllactose- N-neohexaose, sialyllactose-N-fucohexaose II, disialyllactose-N-fucopentaose II, monofucosyldisialyllactose-N-tetraose, Neu5Ac-a2,3-Gal-b1,4-GlcNAc-b1,3-Gal, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-KDO-lactose, 3'-KDO-lactosamine, 3'-KDO-6' sialyllactose, 3'KDO-8-sialyllactose, KDO-2,3Galβ-1,3GalNacβ-1,3Galα-1,4Galβ-1,4Gal, KDO-2,3Galβ-1,3GlcNacβ-1, 3Galβ-1,4Glc, KDO-2,3Galβ-1,4GlcNacβ-1,3Galβ-1,4Glc, 3'-KDO-3-fucosyllactose, Neu5Ac-a2,8-Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-sialyl-2'-fucosyllactose, 6'-sialyl-2'-fucosyllactose, 6'-sialyl-3-fucosyllactose, 3'-sialyl-3-fucosyllactose, Neu5Ac-a2,6-(Neu5Ac-a2,3-)Gal-b1,4-Glc, 3'-sialyl-3-fucosyllactosamine, Fuc-a 1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc, 6'-sialyllactose-N-disaccharide, 3'-sialyllactose-N-disaccharide, Neu5Ac-a2,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,3-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1, 3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Ga l-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-Gal-b1,3-GlcNAc-b1 ,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc-b1,3-G al-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-(Fuc-a1,2-) Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal -b1,4-Glc and any combination thereof; preferably wherein the neutral (uncharged) oligosaccharide containing N-acetylglucosamine is selected from lactose-N-triose II (LN3), lactose-N-tetraose (LNT), lactose-N-neotetraose (LNnT), 6'-galactosyl lactose, 4'-galactosyl lactose, 3'-galactosyl lactose, GlcNAc-b1,6-Gal-b1,4-Glc, lactose-N-hexaose (LNH), lactose-N-neohexaose (LNnH), p-lactose-N-hexaose (pLNH), p-lactose-N-neohexaose (pLNnH), GlcNAc-b1,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, lactose-N-hexaose (LNH), lactose-N-neohexaose (LNnH), p-lactose-N-hexaose (pLNH), p-lactose-N-neohexaose (pLNnH), GlcNAc-b1,6-(GlcNAc-b1,3-)Gal-b1,4-Glc4-Glc, lactose-N-pentaose (LN5), lactose-N-neopentaose, p-lactose-N-pentaose, p-lactose-N-neopentaose, lactose-N-novopentaose I, lactose-N-heptaose (LN7), lactose-N-neoheptaose, p-lactose-N-neoheptaose, p-lactose-N-heptaose, lactose-N-octaose (LNO), lactose-N-neooctaose, allolactose-N-octaose, p-lactose-N-octaose, allolactose-N-neooctaose, novel lactose-N-neooctaose, p-lactose-N-neooctaose (pLNnO), allolactose-N-nonaose, novel lactose-N-nonaose, lactose-N-nonaose (LN9), lactose-N-decaose, allolactose-N-decaose, novel lactose-N-decaose, lactose-N-neodecaose, p-lactose Glyco-N-neodecaose (pLNnD), α1,3-galactosyl lactose-N-neotetraose, GlcNAc-b1,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, GlcNAc-b1,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc and GlcNAc-b1,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc; preferably wherein the milk oligosaccharide is a mammalian milk oligosaccharide (MMO), more preferably a human milk oligosaccharide (HMO); preferably wherein the animal oligosaccharide is selected from N-glycans and O-glycans; preferably wherein the plant oligosaccharide is selected from N-glycans and O-glycans.

[0318] More specifically, the present invention relates to the following preferred embodiments:

[0319] 1. A method for purifying oligosaccharides from a solution, wherein the solution containing the oligosaccharides is a solution selected from the group consisting of a biocatalytic reaction solution, a chemical synthesis solution, a cell culture, and a fermentation product, wherein the oligosaccharides are produced by the biocatalytic reaction solution, the chemical synthesis solution, or by cells cultured in the cell culture or the fermentation, the method comprising subjecting the solution to a stream containing H + a first cation exchange with a cation exchange resin in the form of a cation exchange resin, thereby converting the salt in the solution completely or partially into an acid,

[0320] characterised in that said solution comprising said salt converted completely or partially into acid is passed through a + , K + , Ca 2 + Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + a second cation exchange resin in the form of a cation exchange resin, and wherein:

[0321] - Make it contain H + The eluate obtained in the first cation exchange of the cation exchange resin in the form of + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + the second cation exchange resin in the form of a cation exchange resin, or

[0322] - the method further comprises any one or more processes selected from the list consisting of homogenization, clarification, clearing, concentration, centrifugation, decantation, dilution, pH adjustment in the range of 2 to 5, temperature adjustment, filtration, ultrafiltration, microfiltration, diafiltration, reverse osmosis, nanofiltration, dialysis, use of activated carbon or carbon, use of a solvent in the range of pH 2 to 5, use of an alcohol, use of an aqueous alcohol mixture, use of charcoal, tangential flow high performance filtration, tangential flow ultrafiltration, affinity chromatography, hydrophobic interaction chromatography, gel filtration, ligand exchange chromatography, column chromatography, cation exchange adsorption resin, use of an adsorbent material, evaporation, vacuum evaporation, wiped film evaporation, falling film evaporation, pasteurization, enzymatic treatment, decolorization, drying, and dissolution in a liquid in the range of pH 2 to 5, and

[0323] wherein any one or more of the processes comprises H + After the first cation exchange of the cation exchange resin in the form of Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + Form a cation exchange resin before the second cation exchange.

[0324] 2. The method according to preferred embodiment 1, wherein the cation exchange resin in the first and / or second cation exchange:

[0325] - selected from the list consisting of weak acid cation (WAC) exchange resins and strong acid cation (SAC) exchange resins,

[0326] - having an acrylic-based, methacrylic-based, styrene-based or polystyrene-based matrix, and / or

[0327] - having an acrylic-based, methacrylic-based, styrene-based or polystyrene-based matrix, wherein the matrix further comprises divinylbenzene (DVB).

[0328] 3. The method according to any one of the preferred embodiments 1 or 2, wherein:

[0329] - a flow rate through the first and / or second cation exchange is at least 0.5, at least 1, at least 2, at least 2.5 and / or at least 3 bed volumes / hour (BV / h),

[0330] - the first and / or second cation exchange is performed at a temperature ranging from 0°C to 80°C, 4°C to 60°C, 4°C to 40°C, 10°C to 37°C, 20°C to 30°C, 20°C to 25°C, 22°C to 24°C and / or 23°C to 24°C, and / or

[0331] - the first and / or second cation exchange is performed at a pH in the range of 2 to 5, 3 to 5 and / or 4 to 5.

[0332] 4. The method according to any one of the preceding preferred embodiments, wherein the method further comprises any one or more processes selected from the list consisting of: homogenization, clarification, clearing, concentration, centrifugation, decantation, dilution, pH adjustment, temperature adjustment, filtration, ultrafiltration, microfiltration, diafiltration, reverse osmosis, electrodialysis, electrodeionization, nanofiltration, dialysis, use of activated carbon or carbon, use of a solvent, use of an alcohol, use of an aqueous alcohol mixture, use of charcoal, tangential flow high performance filtration, tangential flow ultrafiltration, affinity chromatography, ion exchange, ion exchange chromatography, mixed bed ion exchange, hydrophobic interaction chromatography, gel filtration, ligand exchange chromatography, column chromatography, cation exchange adsorption resins, anion exchange adsorption resins, use of an adsorbent material, use of an ion exchange resin, evaporation, vacuum evaporation, wiped film evaporation, falling film evaporation, pasteurization, enzymatic treatment, decolorization and drying, and wherein any one or more of the processes:

[0333] (a) In the case of H + Form a cation exchange resin before the first cation exchange,

[0334] (b) any one or more of the processes described herein in the presence of Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + After the second cation exchange of the form cation exchange resin, and / or

[0335] (c) is performed more than once during the process.

[0336] 5. The method according to any one of the preceding preferred embodiments, wherein the temperature of the solution is adjusted to a temperature of:

[0337] - from 36°C to 65°C, from 36°C to 60°C, from 40°C to 55°C and / or from 40°C to 45°C, wherein the temperature is within 5°C of the temperature at which the solution exhibits maximum turbidity, and / or

[0338] - a temperature of from 0°C to 122°C, from 2°C to 80°C, from 4°C to 60°C, from 10°C to 55°C, from 20°C to 45°C, from 21°C to 40°C, from 22°C to 37°C and / or from 25°C to 30°C.

[0339] 6. The method according to any one of the preferred embodiments 4 or 5, wherein the method:

[0340] - consists of two consecutive nanofiltration steps,

[0341] - involves two consecutive electrodialysis steps or no electrodialysis,

[0342] - comprises two consecutive ultrafiltration steps, wherein the molecular weight cut-off of the membrane used in the first ultrafiltration step is higher than the molecular weight cut-off of the membrane used in the second ultrafiltration step,

[0343] - includes clarification by any one or more of microfiltration, centrifugation, flocculation or ultrafiltration,

[0344] - comprising drying selected from the group consisting of spray drying, freeze drying, spray freeze drying, freeze spray drying, belt drying, conveyor belt drying, vacuum belt drying, vacuum conveyor belt drying, drum drying, roller drying, vacuum drum drying, vacuum roller drying and stirred film drying,

[0345] - including filtration using filter aids and / or flocculants,

[0346] - comprising filtration performed using a filter aid, wherein the filter aid is an adsorbent,

[0347] - comprising filtration by using a filter aid, wherein the filter aid is activated carbon,

[0348] - comprising ultrafiltration, wherein the ultrafiltration has a molecular weight cut-off equal to or higher than 1 kDa, 2 kDa, 3 kDa, 4 kDa, 5 kDa, 6 kDa, 7 kDa, 8 kDa, 9 kDa, 10 kDa, 11 kDa, 12 kDa, 13 kDa, 14 kDa, 15 kDa,

[0349] - including nanofiltration, wherein the nanofiltration membrane used in the nanofiltration has ≤ The size exclusion limit,

[0350] - comprising diafiltration of the solution until the conductivity reaches ≤ 15 mS / cm, ≤ 10 mS / cm, ≤ 5 mS / cm, ≤ 1 mS / cm, ≤ 0.1 mS / cm, ≤ 0.01 mS / cm and / or ≤ 0.001 mS / cm,

[0351] - comprising an enzymatic treatment comprising incubating the solution with one or more enzymes selected from the list consisting of glycosidase, lactase, β-galactosidase, fucosidase, sialidase, maltase, amylase, hexamidinase, glucuronidase, trehalase and invertase,

[0352] - enzymatic treatment comprising the conversion of lactose, sucrose, malto-oligosaccharides, maltotriose, sorbitol, trehalose, starch, cellulose, hemicellulose, lignocellulose, molasses, corn syrup and / or high fructose syrup into monosaccharides, and / or

[0353] - is a batch or continuous process.

[0354] 7. The method according to any one of the aforementioned preferred embodiments, wherein the oligosaccharide is selected from the list consisting of: fucosylated oligosaccharides; neutral (uncharged) oligosaccharides; negatively charged oligosaccharides; sialylated oligosaccharides; Lewis-type antigens; oligosaccharides containing N-acetylglucosamine; neutral (uncharged) oligosaccharides containing N-acetylglucosamine; oligosaccharides containing N-acetyllactosamine; oligosaccharides containing lactose-N-disaccharide; oligosaccharides containing galactose; non-fucosylated neutral (uncharged) oligosaccharides; chitosan; oligosaccharides containing chitosan; heparin precursors; chondroitin sulfate; glycosaminoglycan oligosaccharides; heparin; heparan sulfate; dermatan sulfate; hyaluronan or hyaluronic acid; keratan sulfate; milk oligosaccharides; O-antigens; enterobacterial common antigens (ECA); oligosaccharide repeating units present in capsular polysaccharides; peptidoglycans; amino sugars; oligosaccharides of the human ABO blood group system Antigen; Animal oligosaccharide; Plant oligosaccharide; Glucopyranosyl sucrose (Glc-a1,4-Glc-a1,2-Fru); Lactulose-N-triose II (GlcNAc-b1,3-Gal-b1,4-Fru); Lactulose-N-tetraose; Lactulose-N-neotetraose; Uridine diphosphate galactose; Fucosylated oligosaccharide selected from the following list: 2'-fucosyllactose (2'FL ), 3-fucosyllactose (3FL), 4-fucosyllactose (4FL), 6-fucosyllactose (6FL), 2',3-difucosyllactose (diFL), lactose-N-fucopentaose I (LNFPI), Gal-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (Gal-LNFP I), GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (GalNAc-LNFP I), lactose-N-neofucopentaose I (LNnFP I), lactose-N-fucopentaose II (LNFP II), lactose-N-fucopentaose III (LNFP III), lactose-N-fucopentaose V (LNFP V), lactose-N-fucopentaose VI (LNFP VI), lactose-N-neofucopentaose V, lactose-N-difucohexaose I (LNDFH-I), lactose-N-difucohexaose II (LNDFH-II), Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,4-(Fuc-a1,2-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, monofucosyllactose-N-hexaose-III, difucosyllactose-N-hexaose (a), difucosyllactose-N-hexaose, difucosyllactose-N-neohexaose, trifucosyllactose-N-hexaose, a1,3-galactosyl-3-fucosyllactose sugar, Gal-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, 2-fucosyllactulose, 3-fucosyl-N-acetyllactosamine, 2'-fucosyl-N-acetyllactosamine, difucosyl-N-acetyllactosamine, 4-fucosyllactose-N-disaccharide, 2'-fucosyllactose-N-disaccharide, difucosyllactose-N-disaccharide and GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc; sialylated oligosaccharides selected from 3' sialyllactose (3' SL), 6'-sialyllactose (6'SL), 8'-sialyllactose (8'SL), 3,6-disialyllactose (Neu5Ac-α2,3-(Neu5Ac-α2,6)-Gal-β1,4-Glc), 6,6'-disialyllactose (Neu5Ac-α2,6-Gal-β1,4-(Neu5Ac-α2,6)-Glc), 8,3-disialyllactose (Neu5Ac-α2,8-Neu5Ac-α2,3-Gal-β1,4-Glc), 6'-sialyllactosamine, 3'-sialyllactosamine, sialyllactose-N-tetraose a (LSTa), sialyllactose-N-tetraose b (LSTb), sialyllactose-N-tetraose Saccharide c (LSTc), sialyllactose-N-tetraose d (LSTd), disialyllactose-N-tetraose, disialyllactose-N-neotetraose, monosialyllactose-N-hexaose, disialyllactose-N-hexaose I, disialyllactose-N-hexaose II, monosialyllactose-N-neohexaose I, monosialyllactose-N-neohexaose II, disialyllactose-N-neohexaose, 3'-sialyl-3-fucosyllactose, fucodisialyllactose-N-hexaose, disialylmonofucosyllactose-N-neohexaose, sialyllactose-N-fucohexaose II, disialyllactose-N-fucopentaose II, monofucosyldisialyllactose-N-tetraose, Neu5Ac-a2,3-Gal-b1,4-GlcNAc-b1,3-Gal, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-KDO-lactose, 3'-KDO-lactosamine, 3'-KDO-6' sialyllactose, 3'KDO-8-sialyllactose, KDO-2,3Galβ-1,3GalNacβ-1,3Galα-1,4Galβ-1,4Gal, KDO-2,3Galβ-1,3GlcNacβ-1,3Galβ-1,4Glc, KDO-2,3Galβ-1,4GlcNacβ-1,3Galβ-1,4Glc, 3'-KDO-3-fucosyllactose, Neu5Ac- a2,8-Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-sialyl-2'-fucosyllactose, 6'-sialyl-2'-fucosyllactose, 6'-sialyl-3-fucosyllactose, 3'-sialyl-3-fucosyllactose, Neu5Ac-a2,6-(Neu5Ac-a2,3-)Gal-b1,4-Glc, 3'-sialyl-3-fucosyllactosamine, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc, 6'-sialyl-2'-fucosyllactose, 3'-sialyl-3-fucosyllactose, Neu5Ac-a2,6-(G lcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,3-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1 ,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-GlcNAc-b1,3-)Gal -b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc , Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Fuc-a1,4-(Neu5Ac-a2, 3-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-(Neu5 Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, and any combination thereof; a neutral (uncharged) oligosaccharide containing N-acetylglucosamine selected from lacto-N-triose II (LN3), lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), 6'-galactosyl lactose, 4'-galactosyl lactose, 3'-galactosyl lactose Lactose, GlcNAc-b1,6-Gal-b1,4-Glc, lactose-N-hexaose (LNH), lactose-N-neohexaose (LNnH), p-lactose-N-hexaose (pLNH), p-lactose-N-neohexaose (pLNnH), GlcNAc-b1,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, lactose-N-pentaose (LN5), lactose-N-neopentaose, p-lactose-N-pentaose, p-lactose-N-neopentaose, lactose-N-neopentaose I, lactose-N-heptaose (LN7), lactose-N-neoheptaose, p-lactose-N-neoheptaose, p-lactose-N-heptaose, lactose-N-octaose (LNO), lactose-N-neoheptaose, Allolactose-N-octaose, p-lactose-N-octaose, allolactose-N-neooctaose, novel lactose-N-neooctaose, p-lactose-N-neooctaose (pLNnO), allolactose-N-nonaose, novel lactose-N-nonaose, lactose-N-nonaose (LN9), lactose-N-decaose, allolactose-N-decaose, novel lactose-N-decaose, lactose-N-neodecaose, p-lactose-N-neodecaose (pLNnD), a1,3-galactosyl lactose-N-neotetraose, GlcNAc-b1,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, GlcNAc-b1,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc and GlcNAc-b1,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc; mammalian milk oligosaccharide (MMO); human milk oligosaccharide (HMO); animal oligosaccharide selected from N-glycans and O-glycans; plant oligosaccharide selected from N-glycans and O-glycans; sialylated oligosaccharide having at least one sialic acid group selected from the group consisting of Neu4Ac; Neu5 Ac; Neu4,5Ac2; Neu5,7Ac2; Neu5,8Ac2; Neu5,9Ac2; Neu4,5,9Ac3; Neu5,7,9Ac3; Neu5,8,9Ac3; Neu4,5,7,9Ac4; Neu5,7,8,9Ac4; Neu4,5,7,8,9Ac5; Neu5Gc and 2-keto-3-deoxymannosyl-octanone (KDO). ,

[0355] 8. The method according to any one of the preceding preferred embodiments, wherein:

[0356] Prior to purification by the method described,

[0357] - the purity of the oligosaccharide in the solution is <70%, <60%, <50%, <40%, <30%, <20%, <10% based on total dry solids,

[0358] - the ash content of said solution comprising said oligosaccharides is > 10% based on total dry solids,

[0359] - the ash content of the solution comprising the oligosaccharide is > 10% based on total dry solids, wherein the ash comprises any one or more heavy metals selected from the group consisting of lead, arsenic, cadmium, mercury, zinc, manganese, copper, iron, magnesium and calcium, and / or

[0360] - said solution comprising said oligosaccharide comprises a lead content > 0.1 mg / kg dry solid, an arsenic content > 0.2 mg / kg dry solid, a cadmium content > 0.1 mg / kg dry solid and / or a mercury content > 0.5 mg / kg dry solid.

[0361] 9. The method according to any one of the aforementioned preferred embodiments, wherein the oligosaccharide is accompanied by

[0362] - sialic acid,

[0363] - ash content,

[0364] - ash selected from the list consisting of sulfate, phosphate, sodium, chloride, potassium, heavy metals, preferably ammonium, lead, arsenic, cadmium and / or mercury,

[0365] - one or more monosaccharides,

[0366] - one or more activated monosaccharides,

[0367] - one or more phosphorylated monosaccharides,

[0368] - one or more disaccharides, and / or

[0369] - one or more other oligosaccharides selected from the group consisting of: neutral (uncharged) oligosaccharides; negatively charged oligosaccharides; milk oligosaccharides; mammalian milk oligosaccharides (MMO); human milk oligosaccharides (HMO); O-antigens; enterobacterial common antigens (ECA); oligosaccharide repeating units present in capsular polysaccharides; peptidoglycans; amino sugars; Lewis-type antigenic oligosaccharides; antigens of the human ABO blood group system; animal oligosaccharides; animal oligosaccharides selected from N-glycans and O-glycans; plant oligosaccharides; plant oligosaccharides selected from N-glycans and O-glycans; glucopyranosylsucrose (Glc-a1,4-Gl c-a1,2-Fru); lactulose-N-triose II (GlcNAc-b1,3-Gal-b1,4-Fru); lactulose-N-tetraose; lactulose-N-neotetraose; uridine diphosphate galactose; fucosylated oligosaccharides; fucosylated oligosaccharides selected from the following list: 2'-fucosyllactose (2'FL), 3-fucosyllactose (3FL), 4-fucosyllactose (4FL), 6-fucosyllactose (6FL), 2',3-difucosyllactose (diFL), lactose-N-fucopentaose I (LNFP I), Gal-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (Gal-LNFPI), GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (GalNAc-LNFP I), lactose-N-neofucopentaose I (LNnFP I), lactose-N-fucopentaose II (LNFP II), lactose-N-fucopentaose III (LNFPIII), lactose-N-fucopentaose V (LNFP V), lactose-N-fucopentaose VI (LNFP VI), lactose-N-neofucopentaose V, lactose-N-difucohexaose I (LNDFH-I), lactose-N-difucohexaose II (LNDFH-II), Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1, 4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc,Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Gl c. Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,4-(Fuc-a1,2-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, monofucosyllactose-N-hexaose-III, difucosyllactose-N-hexaose (a), difucosyl-lactose-N-hexaose, difucosyl-lactose-N-neohexaose, trifucosyllactose-N-hexaose, a1,3-galactosyl-3-fucosyllactose, Gal-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, 2-fucosyllactulose, 3-fucosyl-N-acetyllactosamine , 2'-fucosyl-N-acetyllactosamine, difucosyl-N-acetyllactosamine, 4-fucosyllactose-N-disaccharide, 2'-fucosyllactose-N-disaccharide, difucosyllactose-N-disaccharide and GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc; sialylated oligosaccharides; sialylated oligosaccharides selected from 3' sialyllactose (3'SL), 6' sialyllactose (6'SL), 8' sialyllactose (8'SL), 3,6-disialyllactose (Neu5Ac-α2,3-(Neu5Ac-α2,6)-Gal-β1,4-Glc), 6,6'-disialyllactose (Neu5Ac-α2,6-Gal-β1,4-Glc), β1,4-(Neu5Ac-α2,6)-Glc), 8,3-disialyl lactose (Neu5Ac-α2,8-Neu5Ac-α2,3-Gal-β1,4-Glc), 6'-sialyllactosamine, 3'-sialyllactosamine, sialyllactose-N-tetraose a (LSTa), sialyllactose-N-tetraose b (LSTb), sialyllactose-N-tetraose c (LSTc), sialyllactose-N-tetraose d (LSTd), disialyl lactose-N-tetraose, disialyl lactose-N-neotetraose, monosialyllactose-N-hexaose, disialyl lactose-N-hexaose I, disialyl lactose-N-hexaose II, monosialyllactose-N-neohexaose I, monosialyllactose -N-neohexaose II, disialyl lactose-N-neohexaose, 3'-sialyl-3-fucosyllactose, fucodisialyl lactose-N-hexaose, disialyl monofucosyllactose-N-neohexaose, sialyl lactose-N-fucohexaose II, disialyl lactose-N-fucopentaose II, monofucosyl disialyl lactose-N-tetraose, Neu5Ac-a2,3-Gal-b1,4-GlcNAc-b1,3-Gal, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-KDO-lactose, 3'-KDO-lactosamine, 3'-KDO-6' sialyllactose, 3'KDO-8-sialyllactose, KDO-2,3Galβ-1,3GalNacβ-1,3Galα-1,4Galβ-1,4Gal, KDO-2,3Galβ-1,3GlcNacβ-1,3Galβ-1,4Glc, KDO-2,3Galβ-1,4GlcNacβ-1,3Galβ-1,4Glc, 3'-KDO-3-fucosyllactose, Neu5Ac-a2,8-Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-sialyl-2'-fucosyllactose, 6'-sialyl-2'-fucosyllactose, 6'-sialyl-3-fucosyllactose, 3'-sialyl-3-fucosyllactose 、Neu5Ac-a2,6-(Neu5Ac-a2,3-)Gal-b1,4-Glc、3'-Sialyl-3-fucosyllactosamine、Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc、6'-sialyllactose-N-disaccharide、3'-sialyllactose-N-disaccharide、Neu5Ac-a2,6-(GlcNAc-b1,3-)Gal-b1,4-Glc、Neu5Ac-a2,6-(Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc、Neu5Ac-a2,3-Gal-b1,4-(Fuc-a 1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-Gl cNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4- (Fuc-a1,3-)Glc, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4- Glc, Neu5Ac-a2,6-(Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-) Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1, 4-Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc and any combination thereof; oligosaccharides containing N-acetylglucosamine; oligosaccharides containing N-acetylglucosamine selected from lactose-N-triose II (LN3), lactose-N-tetraose (LNT), lactose-N-neotetraose (LNnT), 6'-galactosyl lactose, 4'-galactosyl lactose, 3'-galactosyl lactose, GlcNAc-b1,6-Gal-b1,4-Glc, lactose-N-hexaose (LNH), lactose-N-neohexaose (LNnH), p-lactose-N-hexaose (pLNH), p-lactose -N-neo...

Claims

1. A method for purifying oligosaccharides from a solution, wherein the solution containing the oligosaccharides is a solution selected from the group consisting of a biocatalytic reaction solution, a chemical synthesis solution, a cell culture, and a fermentation product, wherein the oligosaccharides are produced by the biocatalytic reaction solution, the chemical synthesis solution, or by cells cultured in the cell culture or the fermentation, the method comprising subjecting the solution to a stream containing H + a first cation exchange with a cation exchange resin in the form of a cation exchange resin, thereby converting the salt in the solution completely or partially into an acid, It is characterized by The solution comprising the salt converted completely or partially into the acid is passed through a + , K + , Ca 2+ Mg 2 + 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + a second cation exchange resin in the form of a cation exchange resin, and wherein: - Make it contain H + The eluate obtained in the first cation exchange of the cation exchange resin in the form of + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + the second cation exchange resin in the form of a cation exchange resin, or - the method further comprises any one or more processes selected from the list consisting of homogenization, clarification, clearing, concentration, centrifugation, decantation, dilution, pH adjustment in the range of 2 to 5, temperature adjustment, filtration, ultrafiltration, microfiltration, diafiltration, reverse osmosis, nanofiltration, dialysis, use of activated carbon or carbon, use of a solvent in the range of pH 2 to 5, use of an alcohol, use of an aqueous alcohol mixture, use of charcoal, tangential flow high performance filtration, tangential flow ultrafiltration, affinity chromatography, hydrophobic interaction chromatography, gel filtration, ligand exchange chromatography, column chromatography, cation exchange adsorption resin, use of an adsorbent material, evaporation, vacuum evaporation, wiped film evaporation, falling film evaporation, pasteurization, enzymatic treatment, decolorization, drying, and dissolution in a liquid in the range of pH 2 to 5, and wherein any one or more of the processes comprises H + After the first cation exchange of the cation exchange resin in the form of Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + Form a cation exchange resin before the second cation exchange.

2. The method according to claim 1, wherein the cation exchange resin in the first and / or second cation exchange: - selected from the list consisting of weak acid cation (WAC) exchange resins and strong acid cation (SAC) exchange resins, - having an acrylic-based, methacrylic-based, styrene-based or polystyrene-based matrix, and / or - having an acrylic-based, methacrylic-based, styrene-based or polystyrene-based matrix, wherein the matrix further comprises divinylbenzene (DVB).

3. The method according to any one of claims 1 or 2, wherein: - a flow rate through the first and / or second cation exchange is at least 0.5, at least 1, at least 2, at least 2.5 and / or at least 3 bed volumes / hour (BV / h), - the first and / or second cation exchange is performed at a temperature ranging from 0°C to 80°C, 4°C to 60°C, 4°C to 40°C, 10°C to 37°C, 20°C to 30°C, 20°C to 25°C, 22°C to 24°C and / or 23°C to 24°C, and / or - the first and / or second cation exchange is performed at a pH in the range of 2 to 5, 3 to 5 and / or 4 to 5.

4. The method according to any one of the preceding claims, wherein the method further comprises any one or more processes selected from the list consisting of: homogenization, clarification, clearing, concentration, centrifugation, decantation, dilution, pH adjustment, temperature adjustment, filtration, ultrafiltration, microfiltration, diafiltration, reverse osmosis, electrodialysis, electrodeionization, nanofiltration, dialysis, using activated carbon or carbon, using a solvent, using an alcohol, using an aqueous alcohol mixture, using charcoal, tangential flow high performance filtration, tangential flow ultrafiltration, affinity chromatography, ion exchange, ion exchange chromatography, mixed bed ion exchange, hydrophobic interaction chromatography, gel filtration, ligand exchange chromatography, column chromatography, cation exchange adsorption resins, anion exchange adsorption resins, using adsorbent materials, using ion exchange resins, evaporation, vacuum evaporation, wiped film evaporation, falling film evaporation, pasteurization, enzymatic treatment, decolorization and drying, and wherein any one or more of the processes: (a) In the case of H + Form a cation exchange resin before the first cation exchange, (b) any one or more of the processes described herein in the presence of Na + , K + , Ca 2+ Mg 2+ 、Fe 2+ 、Fe 3+ 、Al 3+ or NH4 + After the second cation exchange of the form cation exchange resin, and / or (c) is performed more than once during the process.

5. The method according to any one of the preceding claims, wherein the temperature of the solution is adjusted to a temperature of: - from 36°C to 65°C, from 36°C to 60°C, from 40°C to 55°C and / or from 40°C to 45°C, wherein the temperature is within 5°C of the temperature at which the solution exhibits maximum turbidity, and / or - a temperature of from 0°C to 122°C, from 2°C to 80°C, from 4°C to 60°C, from 10°C to 55°C, from 20°C to 45°C, from 21°C to 40°C, from 22°C to 37°C and / or from 25°C to 30°C.

6. The method according to any one of claims 4 or 5, wherein the method: - consists of two consecutive nanofiltration steps, - involves two consecutive electrodialysis steps or no electrodialysis, - comprises two consecutive ultrafiltration steps, wherein the molecular weight cut-off of the membrane used in the first ultrafiltration step is higher than the molecular weight cut-off of the membrane used in the second ultrafiltration step, - includes clarification by any one or more of microfiltration, centrifugation, flocculation or ultrafiltration, - comprising drying selected from the group consisting of spray drying, freeze drying, spray freeze drying, freeze spray drying, belt drying, conveyor belt drying, vacuum belt drying, vacuum conveyor belt drying, drum drying, roller drying, vacuum drum drying, vacuum roller drying and stirred film drying, - including filtration using filter aids and / or flocculants, - comprising filtration performed using a filter aid, wherein the filter aid is an adsorbent, - comprising filtration by using a filter aid, wherein the filter aid is activated carbon, - comprising ultrafiltration, wherein the ultrafiltration has a molecular weight cut-off equal to or higher than 1 kDa, 2 kDa, 3 kDa, 4 kDa, 5 kDa, 6 kDa, 7 kDa, 8 kDa, 9 kDa, 10 kDa, 11 kDa, 12 kDa, 13 kDa, 14 kDa, 15 kDa, - comprising nanofiltration, wherein the nanofiltration membrane used in said nanofiltration has The size exclusion limit, - comprising diafiltration of the solution until the conductivity reaches ≤ 15 mS / cm, ≤ 10 mS / cm, ≤ 5 mS / cm, ≤ 1 mS / cm, ≤ 0.1 mS / cm, ≤ 0.01 mS / cm and / or ≤ 0.001 mS / cm, - comprising an enzymatic treatment comprising incubating the solution with one or more enzymes selected from the list consisting of glycosidase, lactase, β-galactosidase, fucosidase, sialidase, maltase, amylase, hexamidinase, glucuronidase, trehalase and invertase, - enzymatic treatment comprising the conversion of lactose, sucrose, malto-oligosaccharides, maltotriose, sorbitol, trehalose, starch, cellulose, hemicellulose, lignocellulose, molasses, corn syrup and / or high fructose syrup into monosaccharides, and / or - is a batch or continuous process.

7. The method according to any one of the preceding claims, wherein the oligosaccharide is selected from the list consisting of: fucosylated oligosaccharides; neutral (uncharged) oligosaccharides; negatively charged oligosaccharides; sialylated oligosaccharides; Lewis-type antigens; oligosaccharides containing N-acetylglucosamine; neutral (uncharged) oligosaccharides containing N-acetylglucosamine; oligosaccharides containing N-acetyllactosamine; oligosaccharides containing lactose-N-disaccharide; oligosaccharides containing galactose; non-fucosylated neutral (uncharged) oligosaccharides; chitosan; oligosaccharides containing chitosan; heparin precursors; chondroitin sulfate; glycosaminoglycan oligosaccharides; heparin; heparan sulfate; dermatan sulfate; hyaluronan or hyaluronic acid; keratan sulfate; milk oligosaccharides; O-antigens; enterobacterial common antigen (ECA); Oligosaccharide repeating units in capsular polysaccharides; peptidoglycan; amino sugars; antigens of the human ABO blood group system; animal oligosaccharides; plant oligosaccharides; glucopyranosylsucrose (Glc-a1,4-Glc-a1,2-Fru); lactulose-N-triose II (GlcNAc-b1,3-Gal-b1,4-Fru); lactulose-N-tetraose; lactulose-N-neotetraose; uridine diphosphate galactose; fucosylated oligosaccharides selected from the following list: 2'-fucosyllactose (2'FL), 3-fucosyllactose (3FL), 4-fucosyllactose (4FL), 6-fucosyllactose (6FL), 2',3-difucosyllactose (diFL), lactose-N-fucopentaose I (LNFP I), Gal-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (Gal-LNFP I), GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (GalNAc-LNFP I), lactose-N-neofucopentaose I (LNnFPI), lactose-N-fucopentaose II (LNFP II), lactose-N-fucopentaose III (LNFP III), lactose-N-fucopentaose V (LNFP V), lactose-N-fucopentaose VI (LNFP VI), lactose-N-neofucopentaose V, lactose-N-difucohexaose I (LNDFH-I), lactose-N-difucohexaose II (LNDFH-II), Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,4-(Fuc-a1,2-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, monofucosyllactose-N-hexaose-III, difucosyllactose-N-hexaose (a), difucosyllactose-N-hexaose, difucosyllactose-N-neohexaose, trifucosyllactose-N-hexaose, a1,3-galactosyl-3-fucosyllactose sugar, Gal-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, 2-fucosyllactulose, 3-fucosyl-N-acetyllactosamine, 2'-fucosyl-N-acetyllactosamine, difucosyl-N-acetyllactosamine, 4-fucosyllactose-N-disaccharide, 2'-fucosyllactose-N-disaccharide, difucosyllactose-N-disaccharide and GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc; sialylated oligosaccharides selected from 3' sialyllactose (3' SL), 6'-sialyllactose (6'SL), 8'-sialyllactose (8'SL), 3,6-disialyllactose (Neu5Ac-α2,3-(Neu5Ac-α2,6)-Gal-β1,4-Glc), 6,6'-disialyllactose (Neu5Ac-α2,6-Gal-β1,4-(Neu5Ac-α2,6)-Glc), 8,3-disialyllactose (Neu5Ac-α2,8-Neu5Ac-α2,3-Gal-β1,4-Glc), 6'-sialyllactosamine, 3'-sialyllactosamine, sialyllactose-N-tetraose a (LSTa), sialyllactose-N-tetraose b (LSTb), sialyllactose-N-tetraose Saccharide c (LSTc), sialyllactose-N-tetraose d (LSTd), disialyllactose-N-tetraose, disialyllactose-N-neotetraose, monosialyllactose-N-hexaose, disialyllactose-N-hexaose I, disialyllactose-N-hexaose II, monosialyllactose-N-neohexaose I, monosialyllactose-N-neohexaose II, disialyllactose-N-neohexaose, 3'-sialyl-3-fucosyllactose, fucodisialyllactose-N-hexaose, disialylmonofucosyllactose-N-neohexaose, sialyllactose-N-fucohexaose II, disialyllactose-N-fucopentaose II, monofucosyldisialyllactose-N-tetraose, Neu5Ac-a2,3-Gal-b1,4-GlcNAc-b1,3-Gal, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-KDO-lactose, 3'-KDO-lactosamine, 3'-KDO-6' sialyllactose, 3'KDO-8-sialyllactose, KDO-2,3Galβ-1,3GalNacβ-1,3Galα-1,4Galβ-1,4Gal, KDO-2,3Galβ-1,3GlcNacβ-1,3Galβ-1,4Glc, KDO-2,3Galβ-1,4GlcNacβ-1,3Galβ-1,4Glc, 3'-KDO-3-fucosyllactose, Neu5Ac- a2,8-Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-sialyl-2'-fucosyllactose, 6'-sialyl-2'-fucosyllactose, 6'-sialyl-3-fucosyllactose, 3'-sialyl-3-fucosyllactose, Neu5Ac-a2,6-(Neu5Ac-a2,3-)Gal-b1,4-Glc, 3'-sialyl-3-fucosyllactosamine, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc, 6'-sialyl-2'-fucosyllactose, 3'-sialyl-3-fucosyllactose, Neu5Ac-a2,6-(G lcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,3-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1 ,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-GlcNAc-b1,3-)Gal -b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc , Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Fuc-a1,4-(Neu5Ac-a2, 3-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-(Neu5 Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, and any combination thereof; a neutral (uncharged) oligosaccharide containing N-acetylglucosamine selected from lacto-N-triose II (LN3), lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), 6'-galactosyl lactose, 4'-galactosyl lactose, 3'-galactosyl lactose Lactose, GlcNAc-b1,6-Gal-b1,4-Glc, lactose-N-hexaose (LNH), lactose-N-neohexaose (LNnH), p-lactose-N-hexaose (pLNH), p-lactose-N-neohexaose (pLNnH), GlcNAc-b1,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, lactose-N-pentaose (LN5), lactose-N-neopentaose, p-lactose-N-pentaose, p-lactose-N-neopentaose, lactose-N-neopentaose I, lactose-N-heptaose (LN7), lactose-N-neoheptaose, p-lactose-N-neoheptaose, p-lactose-N-heptaose, lactose-N-octaose (LNO), lactose-N-neoheptaose, Allolactose-N-octaose, p-lactose-N-octaose, allolactose-N-neooctaose, novel lactose-N-neooctaose, p-lactose-N-neooctaose (pLNnO), allolactose-N-nonaose, novel lactose-N-nonaose, lactose-N-nonaose (LN9), lactose-N-decaose, allolactose-N-decaose, novel lactose-N-decaose, lactose-N-neodecaose, p-lactose-N-neodecaose (pLNnD), a1,3-galactosyl lactose-N-neotetraose, GlcNAc-b1,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, GlcNAc-b1,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc and GlcNAc-b1,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc; mammalian milk oligosaccharide (MMO); human milk oligosaccharide (HMO); animal oligosaccharide selected from N-glycans and O-glycans; plant oligosaccharide selected from N-glycans and O-glycans; sialylated oligosaccharide having at least one sialic acid group selected from the group consisting of Neu4Ac; Neu5 Ac; Neu4,5Ac2; Neu5,7Ac2; Neu5,8Ac2; Neu5,9Ac2; Neu4,5,9Ac3; Neu5,7,9Ac3; Neu5,8,9Ac3; Neu4,5,7,9Ac4; Neu5,7,8,9Ac4; Neu4,5,7,8,9Ac5; Neu5Gc and 2-keto-3-deoxymannosyl-octanone (KDO). , 8. A method according to any one of the preceding claims, wherein: Prior to purification by the method described, - the purity of the oligosaccharide in the solution is <70%, <60%, <50%, <40%, <30%, <20%, <10% based on total dry solids, - the ash content of said solution comprising said oligosaccharides is > 10% based on total dry solids, - the ash content of the solution comprising the oligosaccharide is > 10% based on total dry solids, wherein the ash comprises any one or more heavy metals selected from the group consisting of lead, arsenic, cadmium, mercury, zinc, manganese, copper, iron, magnesium and calcium, and / or - said solution comprising said oligosaccharide comprises a lead content > 0.1 mg / kg dry solid, an arsenic content > 0.2 mg / kg dry solid, a cadmium content > 0.1 mg / kg dry solid and / or a mercury content > 0.5 mg / kg dry solid.

9. The method according to any one of the preceding claims, wherein the oligosaccharide is accompanied in the solution by - sialic acid, - ash content, - ash selected from the list consisting of sulfate, phosphate, sodium, chloride, potassium, heavy metals, preferably ammonium, lead, arsenic, cadmium and / or mercury, - one or more monosaccharides, - one or more activated monosaccharides, - one or more phosphorylated monosaccharides, - one or more disaccharides, and / or one or more other oligosaccharides selected from the group consisting of: neutral (uncharged) oligosaccharides; negatively charged oligosaccharides; milk oligosaccharides; mammalian milk oligosaccharides (MMO); human milk oligosaccharides (HMO); O-antigens; enterobacterial common antigens (ECA); oligosaccharide repeating units present in capsular polysaccharides; peptidoglycans; amino sugars; Lewis-type antigenic oligosaccharides; antigens of the human ABO blood group system; animal oligosaccharides; animal oligosaccharides selected from N-glycans and O-glycans; plant oligosaccharides; plant oligosaccharides selected from N-glycans and O-glycans; glucopyranosylsucrose (Glc-a1,4-Glc-a1,2-Fru); lactulose-N-trisaccharide II (GlcNAc-b1,3-Gal-b1,4-Fru); lactulose-N-tetraose; lactulose-N-neotetraose; uridine diphosphate galactose; fucosylated oligosaccharides;Fucosylated oligosaccharides selected from the group consisting of 2'-fucosyllactose (2'FL), 3-fucosyllactose (3FL), 4-fucosyllactose (4FL), 6-fucosyllactose (6FL), 2',3-difucosyllactose (diFL), lactose-N-fucopentaose I (LNFP I), Gal-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (Gal-LNFP I), GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (GalNAc-LNFP I), lactose-N-neofucopentaose I (LNnFP I), lactose-N-fucopentaose II (LNFP II), and lactose-N-fucopentaose II (LNFP II). II), lactose-N-fucopentaose III (LNFPIII), lactose-N-fucopentaose V (LNFP V), lactose-N-fucopentaose VI (LNFP VI), lactose-N-neofucopentaose V, lactose-N-difucohexaose I (LNDFH-I), lactose-N-difucohexaose II (LNDFH-II), Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1, 4-Glc, Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,4-(Fuc-a1,2-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, monofucosyllactose-N-hexaose-III, difucosyllactose; -N-hexaose (a), difucosyl-lactose-N-hexaose, difucosyl-lactose-N-neohexaose, trifucosyllactose-N-hexaose, a1,3-galactosyl-3-fucosyllactose, Gal-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, 2-fucosyllactulose, 3-fucosyl-N-acetyllactosamine, 2'-fucosyl-N-acetyllactosamine, difucosyl-N-acetyllactosamine, 4-fucosyllactose-N-disaccharide, 2'-fucosyllactose-N-disaccharide, di Fucosyllactose-N-disaccharide and GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc; sialylated oligosaccharides; sialylated oligosaccharides selected from 3' sialyllactose (3'SL), 6' sialyllactose (6'SL), 8' sialyllactose (8'SL), 3,6-disialyllactose (Neu5Ac-α2,3-(Neu5Ac-α2,6)-Gal-β1,4-Glc), 6,6'-disialyllactose (Neu5Ac-α2,6-Gal-β1,4-(Neu5Ac-α2,6)-Glc), 8,3-disialyllactose (Neu5Ac-α2,8-Neu5Ac-α2,3-Gal-β 1,4-Glc), 6'-sialyllactosamine, 3'-sialyllactosamine, sialyllactose-N-tetraose a (LSTa), sialyllactose-N-tetraose b (LSTb), sialyllactose-N-tetraose c (LSTc), sialyllactose-N-tetraose d (LSTd), disialyllactose-N-tetraose, disialyllactose-N-neotetraose, monosialyllactose-N-hexaose, disialyllactose-N-hexaose I, disialyllactose-N-hexaose II, monosialyllactose-N-neohexaose I, monosialyllactose-N-neohexaose II, disialyllactose-N-neohexaose, 3'-sialyl-3-fucosyllactose, fucodisialyllactose-N-hexaose, disialylmonofucosyllactose -N-neohexaose, sialyllactose-N-fucohexaose II, disialyllactose-N-fucopentaose II, monofucosyldisialyllactose-N-tetraose, Neu5Ac-a2,3-Gal-b1,4-GlcNAc-b1,3-Gal, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-KDO-lactose, 3'-KDO-lactosamine, 3'-KDO-6' sialyllactose, 3'KDO-8-sialyllactose, KDO-2,3Galβ-1,3GalNacβ-1,3Galα-1,4Galβ-1,4Gal, KDO-2,3Galβ-1,3GlcNacβ-1,3Galβ-1,4Glc, KDO-2,3Galβ-1,4GlcNacβ-1,3Galβ-1,4Glc, 3'-KDO-3-fucosyllactose, Neu5Ac-a2,8-Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-sialyl-2'-fucosyllactose, 6'-sialyl-2'-fucosyllactose, 6'-sialyl-3-fucosyllactose, 3'-sialyl-3-fucosyllactose, Neu5Ac-a2,6-(Neu5Ac-a2,3-)Gal-b1,4-Glc, 3'-sialyl-3-fucosyllactosamine, Fuc-a 1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc, 6'-sialyllactose-N-disaccharide, 3'-sialyllactose-N-disaccharide, Neu5Ac-a2,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,3-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1, 4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Ga l-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc -a1,3-)Glc, Neu5Ac-a2,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc , Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-Glc NAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1 ,4-Glc and any combination thereof; oligosaccharides containing N-acetylglucosamine; oligosaccharides containing N-acetylglucosamine selected from lactose-N-triose II (LN3), lactose-N-tetraose (LNT), lactose-N-neotetraose (LNnT), 6'-galactosyl lactose, 4'-galactosyl lactose, 3'-galactosyl lactose, GlcNAc-b1,6-Gal-b1,4-Glc, lactose-N-hexaose (LNH), lactose-N-neohexaose (LNnH), p-lactose-N-hexaose (pLNH), p-lactose-N-neohexaose (pLNnH), GlcNAc-b1,6-(Glc NAc-b1,3-)Gal-b1,4-Glc, lactose-N-pentaose (LN5), lactose-N-neopentaose, p-lactose-N-pentaose, p-lactose-N-neopentaose, lactose-N-novel pentasaccharide I, lactose-N-heptasaccharide (LN7), lactose-N-neoheptasaccharide, p-lactose-N-neoheptasaccharide, p-lactose-N-heptasaccharide, lactose-N-octaose (LNO), lactose-N-neooctaose, allolactose-N-octaose, p-lactose-N-octaose, allolactose-N-neooctaose, novel lactose-N-neooctaose, p-lactose-N-neooctaose (pLNnO), allolactose-N-nonasaccharide, novel lactose-N-nonasaccharide, lactose Lactose-N-nonasaccharide (LN9), lactose-N-decaose, allolactose-N-decaose, novel lactose-N-decaose, lactose-N-neodecaose, para-lactose-N-neodecaose (pLNnD), a1,3-galactosyllacto-N-neotetraose, GlcNAc-b1,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, GlcNAc-b1,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc and GlcNAc-b1,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc; neutral (uncharged) oligosaccharides containing N-acetylglucosamine; oligosaccharides containing N-acetyllactosamine; oligosaccharides containing lactose-N-disaccharide; non-fucosylated neutral (uncharged) oligosaccharides; chitosan; chitosan-containing oligosaccharides; heparin precursors; chondroitin sulfate; glycosaminoglycan oligosaccharides; heparin; heparan sulfate; dermatan sulfate; hyaluronan; hyaluronic acid; and keratan sulfate.

10. The method according to any one of the preceding claims, wherein the solution is: - using a cell culture of cells or metabolically engineered cells, wherein the oligosaccharide is produced by the cells, the cell culture comprising the oligosaccharide, biomass, culture medium components and contaminants, wherein the purity of the oligosaccharide in the cell culture prior to purification by the method is <70%, <60%, <50%, <40%, <30%, <20%, <10%, - using a cell culture of cells or metabolically engineered cells, wherein said oligosaccharides are produced by said cells, said cell culture comprising said oligosaccharides, biomass, culture medium components and contaminants, wherein when present, biomass separated during said process is optionally recycled to said cell culture, - using a cell culture of at least one cell that has been metabolically engineered to produce said oligosaccharides, and / or - using a cell culture of at least one cell that has been metabolically engineered to produce said oligosaccharide and one or more of: i) sialic acid, ii) one or more monosaccharides, iii) one or more activated monosaccharides, iv) one or more phosphorylated monosaccharides, v) one or more disaccharides and / or vi) one or more other oligosaccharides.

11. The method of any one of the preceding claims, wherein the cell is a bacterium, a fungus, a yeast, a plant cell, an animal cell or a protozoan cell.

12. The method according to any one of the preceding claims, wherein the cells are cultured in the following medium: - a medium comprising a carbon source comprising monosaccharides, disaccharides, oligosaccharides, polysaccharides, polyols, glycerol, a complex medium comprising molasses, corn steep liquor, peptone, tryptone or yeast extract; - a culture medium comprising a carbon source, wherein the carbon source is selected from the group consisting of glucose, N-acetylglucosamine (GlcNAc), glycerol, fructose, sucrose, maltose, lactose, arabinose, maltooligosaccharides, maltotriose, sorbitol, xylose, rhamnose, galactose, mannose, methanol, ethanol, trehalose, starch, cellulose, hemicellulose, molasses, corn syrup, high fructose syrup, acetate, citrate, lactate and pyruvate; - chemically defined medium, and / or - Minimal salts medium comprising sulfate, phosphate, chloride, ammonium, calcium, magnesium, sodium, potassium, iron, copper, zinc, manganese, cobalt and / or selenium.

13. The method according to any one of the preceding claims, wherein the oligosaccharide is accompanied in the solution by one or more other oligosaccharides, wherein: - at least one of said other oligosaccharides has the same degree of polymerization (DP) as said oligosaccharide, - at least one of said other oligosaccharides has a different DP than said oligosaccharide, - all said other oligosaccharides have the same DP, or - all said other oligosaccharides have a different DP than said oligosaccharide.

14. The method according to any one of claims 9 to 13, wherein: - purifying said oligosaccharides from said sialic acid and / or said ash by said method, or - purifying said oligosaccharide and said one or more other oligosaccharides from said sialic acid and / or said ash by said method.

15. A method according to any one of the preceding claims, wherein: - the purity of the oligosaccharides obtained in the purified oligosaccharide solution at the end of the process is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, based on total dry solids, and / or - a purified oligosaccharide yield of at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99% obtained in the purified oligosaccharide solution at the end of the process.

16. The process according to any one of the preceding claims, wherein the purified oligosaccharide solution obtained at the end of the process: - has an ash content of ≤10%, ≤9%, ≤8%, ≤7%, ≤6%, ≤5%, ≤4%, ≤3%, ≤2%, ≤1% and / or ≤0.5%, based on total dry solids, - having an ash content of ≤ 10% based on total dry solids, wherein the ash contains any one or more heavy metals selected from the group consisting of lead, arsenic, cadmium, mercury, zinc, manganese, copper, iron, magnesium and calcium, - have an ash content of ≤ 10% based on total dry solids and a lead content of less than 0.1 mg / kg dry solids, less than 0.02 mg / kg dry solids and / or less than 0.01 mg / kg dry solids, - have an ash content of ≤ 10% based on total dry solids and an arsenic content of less than 0.2 mg / kg dry solids, less than 0.05 mg / kg dry solids and / or less than 0.02 mg / kg dry solids, - have an ash content of ≤ 10% based on total dry solids and a cadmium content of less than 0.1 mg / kg dry solids and / or less than 0.01 mg / kg dry solids, - have an ash content of ≤ 10% based on total dry solids and a mercury content of less than 0.5 mg / kg dry solids, less than 0.1 mg / kg dry solids and / or less than 0.005 mg / kg dry solids, - is filter sterilized and / or subjected to endotoxin removal, and / or subjected to endotoxin removal by filtration through a 3 kDa filter, - having a protein content of less than 100 mg / kg dry solids, a DNA content of less than 10 ng / g dry solids and / or an endotoxin content of less than 10,000 EU / g dry solids, - does not contain DNA, recombinant DNA, proteins, proteins derived from recombinant microorganisms and / or recombinant genetic material, - having less than 125 μg / kg of 3-MCPD, wherein the 3-MCPD is the sum of 3-monochloropropanediol (3-MCPD) and 3-MCPD fatty acid esters, - having less than 0.5 μg / kg epichlorohydrin, - further i) concentrated to a syrup of at least 20% dry matter, at least 30% dry matter, at least 40% dry matter; ii) crystallized; iii) dried to a powder or iv) granulated, - further concentrated to an oligosaccharide concentration of >100 g / L, >200 g / L, >300 g / L, >400 g / L, >500 g / L, >600 g / L and / or 300 g / L to 650 g / L, and / or at a temperature of <80°C, <60°C, <50°C, 20°C to 50°C and / or 30°C to 45°C, - further concentration by processes including the use of vacuum evaporation or reverse osmosis or nanofiltration, - comprising oligosaccharides concentrated to a concentration > 1.5 M and cooled to a temperature < 25°C, and / or < 8°C, to obtain a crystalline material of oligosaccharides, - drying by any one or more drying steps selected from the group consisting of spray drying, freeze drying, evaporation, precipitation, spray freeze drying, freeze spray drying, belt drying, conveyor belt drying, vacuum belt drying, vacuum conveyor belt drying, drum drying, roller drying, vacuum drum drying, vacuum roller drying and stirred film drying, - drying by spray drying, freeze drying or stirred film drying and wherein the pH of the purified oligosaccharide solution is in the range of 2 to 7, 3 to 6 and / or 4 to 5, and / or - Spray drying at a nozzle temperature of 110-150°C, 120-140°C and / or 125-135°C and / or an outlet temperature of 60-80°C and / or 65-70°C at an oligosaccharide solution concentration of 20-60 (w / v), 30-50 (w / v) and / or 35-45 (w / v).

17. A purified oligosaccharide solution, purified oligosaccharide or purified oligosaccharide mixture obtainable or obtained by a process according to any one of the preceding claims.

18. Purified oligosaccharide obtainable or obtained by the process according to any one of claims 1 to 16, wherein the purified oligosaccharide solution comprising the purified oligosaccharide is i) dried, spray dried or dried by a stirred thin film dryer; ii) freeze dried, iii) crystallized or iv) concentrated to a syrup of at least 20% dry matter, at least 30% dry matter or at least 40% dry matter.

19. Oligosaccharides purified according to the method of any one of claims 1 to 16, and wherein the purified oligosaccharides obtained after said method: - have an ash content of ≤10% based on total dry solids and a lead content of less than 0.02 mg / kg dry solids or less than 0.01 mg / kg dry solids, - have an ash content of ≤ 10% based on total dry solids and an arsenic content of less than 0.05 mg / kg dry solids or less than 0.02 mg / kg dry solids, - have an ash content of ≤10% based on total dry solids and a cadmium content of less than 0.01 mg / kg dry solids, - have an ash content of ≤ 10% based on total dry solids and a mercury content of less than 0.1 mg / kg dry solids or less than 0.005 mg / kg dry solids, - does not contain DNA, recombinant DNA, proteins, proteins derived from recombinant microorganisms and / or recombinant genetic material, - has less than 125 μg / kg of 3-MCPD, wherein the 3-MCPD is the sum of 3-monochloropropanediol (3-MCPD) and 3-MCPD fatty acid esters, and / or - Having less than 0.5 μg / kg epichlorohydrin.

20. A dry powder of a purified oligosaccharide solution obtained from the process according to claim 16, wherein the dry powder: - contains ≤15 wt. %, ≤10 wt. %, ≤7 wt. % and / or ≤5 wt. % water, - has an average particle size of 50 to 250 μm, 95 to 120 μm or 110 to 120 μm, wherein the particle size is determined by laser diffraction, - have an ash content of ≤10% based on total dry solids and a lead content of less than 0.02 mg / kg dry solids or less than 0.01 mg / kg dry solids, - have an ash content of ≤ 10% based on total dry solids and an arsenic content of less than 0.05 mg / kg dry solids or less than 0.02 mg / kg dry solids, - have an ash content of ≤10% based on total dry solids and a cadmium content of less than 0.01 mg / kg dry solids, - have an ash content of ≤ 10% based on total dry solids and a mercury content of less than 0.1 mg / kg dry solids or less than 0.005 mg / kg dry solids, - does not contain DNA, recombinant DNA, proteins, proteins derived from recombinant microorganisms and / or recombinant genetic material, - has less than 125 μg / kg of 3-MCPD, wherein the 3-MCPD is the sum of 3-monochloropropanediol (3-MCPD) and 3-MCPD fatty acid esters, and / or - Having less than 0.5 μg / kg epichlorohydrin.

21. A purified oligosaccharide comprising an ash content of ≤ 10% based on total dry solids, wherein the ash comprises: -Less than 0.02 mg / kg dry solid or less than 0.01 mg / kg dry solid lead content, - an arsenic content of less than 0.05 mg / kg dry solid or less than 0.02 mg / kg dry solid, - a cadmium content of less than 0.01 mg / kg dry solids, and / or - Mercury content less than 0.1 mg / kg dry solid or less than 0.005 mg / kg dry solid.

22. The purified oligosaccharide of claim 21, wherein the purified oligosaccharide is selected from the group consisting of: fucosylated oligosaccharides; neutral (uncharged) oligosaccharides; negatively charged oligosaccharides; sialylated oligosaccharides; Lewis-type antigens; oligosaccharides containing N-acetylglucosamine; neutral (uncharged) oligosaccharides containing N-acetylglucosamine; oligosaccharides containing N-acetyllactosamine; oligosaccharides containing lactose-N-disaccharide; oligosaccharides containing galactose; non-fucosylated neutral (uncharged) oligosaccharides; chitosan; oligosaccharides containing chitosan; heparin precursors; chondroitin sulfate; glycosaminoglycan oligosaccharides; heparin; heparan sulfate; dermatan sulfate; hyaluronan or hyaluronic acid; keratan sulfate; milk oligosaccharides; O-antigens; enterobacterial common antigen (ECA); Oligosaccharide repeating units in capsular polysaccharides; peptidoglycan; amino sugars; antigens of the human ABO blood group system; animal oligosaccharides; plant oligosaccharides; glucopyranosylsucrose (Glc-a1,4-Glc-a1,2-Fru); lactulose-N-triose II (GlcNAc-b1,3-Gal-b1,4-Fru); lactulose-N-tetraose; lactulose-N-neotetraose; uridine diphosphate galactose; fucosylated oligosaccharides selected from the following list: 2'-fucosyllactose (2'FL), 3-fucosyllactose (3FL), 4-fucosyllactose (4FL), 6-fucosyllactose (6FL), 2',3-difucosyllactose (diFL), lactose-N-fucopentaose I (LNFP I), Gal-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc(Gal-LNFP I), GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc(GalNAc-LNFP I), Lactose-N-neofucopentaose I (LNnFP I), Lactose-N-fucopentaose II (LNFP II), Lactose-N-fucopentaose III (LNFP III), Lactose-N-fucopentaose V (LNFPV), Lactose-N-fucopentaose VI (LNFP VI), lactose-N-neofucopentaose V, lactose-N-difucohexaose I (LNDFH-I), lactose-N-difucohexaose II (LNDFH-II), Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,4-(Fuc-a1,2-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, monofucosyllactose-N-hexaose-III, difucosyllactose-N-hexaose (a), difucosyllactose-N-hexaose, difucosyllactose-N-neohexaose, trifucosyllactose-N-hexaose, a1,3-galactosyl-3-fucosyllactose sugar, Gal-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, 2-fucosyllactulose, 3-fucosyl-N-acetyllactosamine, 2'-fucosyl-N-acetyllactosamine, difucosyl-N-acetyllactosamine, 4-fucosyllactose-N-disaccharide, 2'-fucosyllactose-N-disaccharide, difucosyllactose-N-disaccharide and GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc; sialylated oligosaccharides selected from 3' sialyllactose (3' SL), 6'-sialyllactose (6'SL), 8'-sialyllactose (8'SL), 3,6-disialyllactose (Neu5Ac-α2,3-(Neu5Ac-α2,6)-Gal-β1,4-Glc), 6,6'-disialyllactose (Neu5Ac-α2,6-Gal-β1,4-(Neu5Ac-α2,6)-Glc), 8,3-disialyllactose (Neu5Ac-α2,8-Neu5Ac-α2,3-Gal-β1,4-Glc), 6'-sialyllactosamine, 3'-sialyllactosamine, sialyllactose-N-tetraose a (LSTa), sialyllactose-N-tetraose b (LSTb), sialyllactose-N-tetraose Saccharide c (LSTc), sialyllactose-N-tetraose d (LSTd), disialyllactose-N-tetraose, disialyllactose-N-neotetraose, monosialyllactose-N-hexaose, disialyllactose-N-hexaose I, disialyllactose-N-hexaose II, monosialyllactose-N-neohexaose I, monosialyllactose-N-neohexaose II, disialyllactose-N-neohexaose, 3'-sialyl-3-fucosyllactose, fucodisialyllactose-N-hexaose, disialylmonofucosyllactose-N-neohexaose, sialyllactose-N-fucohexaose II, disialyllactose-N-fucopentaose II, monofucosyldisialyllactose-N-tetraose, Neu5Ac-a2,3-Gal-b1,4-GlcNAc-b1,3-Gal, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-KDO-lactose, 3'-KDO-lactosamine, 3'-KDO-6' sialyllactose, 3'KDO-8-sialyllactose, KDO-2,3Galβ-1,3GalNacβ-1,3Galα-1,4Galβ-1,4Gal, KDO-2,3Galβ-1,3GlcNacβ-1,3Galβ-1,4Glc, KDO-2,3Galβ-1,4GlcNacβ-1,3Galβ-1,4Glc, 3'-KDO-3-fucosyllactose, Neu5Ac- a2,8-Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-sialyl-2'-fucosyllactose, 6'-sialyl-2'-fucosyllactose, 6'-sialyl-3-fucosyllactose, 3'-sialyl-3-fucosyllactose, Neu5Ac-a2,6-(Neu5Ac-a2,3-)Gal-b1,4-Glc, 3'-sialyl-3-fucosyllactosamine, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc, 6'-sialyl-2'-fucosyllactose, 3'-sialyl-3-fucosyllactose, Neu5Ac-a2,6-(G lcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,3-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1 ,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-GlcNAc-b1,3-)Gal -b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc , Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Fuc-a1,4-(Neu5Ac-a2, 3-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-(Neu5 Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, and any combination thereof; a neutral (uncharged) oligosaccharide containing N-acetylglucosamine selected from lacto-N-triose II (LN3), lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), 6'-galactosyl lactose, 4'-galactosyl lactose, 3'-galactosyl lactose Lactose, GlcNAc-b1,6-Gal-b1,4-Glc, lactose-N-hexaose (LNH), lactose-N-neohexaose (LNnH), p-lactose-N-hexaose (pLNH), p-lactose-N-neohexaose (pLNnH), GlcNAc-b1,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, lactose-N-pentaose (LN5), lactose-N-neopentaose, p-lactose-N-pentaose, p-lactose-N-neopentaose, lactose-N-neopentaose I, lactose-N-heptaose (LN7), lactose-N-neoheptaose, p-lactose-N-neoheptaose, p-lactose-N-heptaose, lactose-N-octaose (LNO), lactose-N-neoheptaose, Allolactose-N-octaose, p-lactose-N-octaose, allolactose-N-neooctaose, novel lactose-N-neooctaose, p-lactose-N-neooctaose (pLNnO), allolactose-N-nonaose, novel lactose-N-nonaose, lactose-N-nonaose (LN9), lactose-N-decaose, allolactose-N-decaose, novel lactose-N-decaose, lactose-N-neodecaose, p-lactose-N-neodecaose (pLNnD), a1,3-galactosyl lactose-N-neotetraose, GlcNAc-b1,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, GlcNAc-b1,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc and GlcNAc-b1,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc; mammalian milk oligosaccharide (MMO); human milk oligosaccharide (HMO); animal oligosaccharide selected from N-glycans and O-glycans; plant oligosaccharide selected from N-glycans and O-glycans.

23. A sugar composition comprising an ash content of ≤10% based on total dry solids and an oligosaccharide content of ≥90% based on total dry solids, wherein the ash comprises: -Less than 0.02 mg / kg dry solid or less than 0.01 mg / kg dry solid lead content, - an arsenic content of less than 0.05 mg / kg dry solid or less than 0.02 mg / kg dry solid, - a cadmium content of less than 0.01 mg / kg dry solids, and / or - Mercury content less than 0.1 mg / kg dry solid or less than 0.005 mg / kg dry solid.

24. The composition of claim 23, wherein the composition comprises more than one oligosaccharide and / or wherein the oligosaccharide is selected from the list consisting of: fucosylated oligosaccharides; neutral (uncharged) oligosaccharides; negatively charged oligosaccharides; sialylated oligosaccharides; Lewis-type antigens; oligosaccharides containing N-acetylglucosamine; neutral (uncharged) oligosaccharides containing N-acetylglucosamine; oligosaccharides containing N-acetyllactosamine; oligosaccharides containing lactose-N-disaccharide; oligosaccharides containing galactose; non-fucosylated neutral (uncharged) oligosaccharides; chitosan; oligosaccharides containing chitosan; heparin precursors; chondroitin sulfate; glycosaminoglycan oligosaccharides; heparin; heparan sulfate; dermatan sulfate; hyaluronan or hyaluronic acid; keratan sulfate; milk oligosaccharides; O-antigens; enterobacterial common antibacterial proto (ECA); oligosaccharide repeating units present in capsular polysaccharides; peptidoglycan; amino sugars; antigens of the human ABO blood group system; animal oligosaccharides; plant oligosaccharides; glucopyranosylsucrose (Glc-a1,4-Glc-a1,2-Fru); lactulose-N-triose II (GlcNAc-b1,3-Gal-b1,4-Fru); lactulose-N-tetraose; lactulose-N-neotetraose; uridine diphosphate galactose; fucosylated oligosaccharides selected from the list comprising 2'-fucosyllactose (2'FL), 3-fucosyllactose (3FL), 4-fucosyllactose (4FL), 6-fucosyllactose (6FL), 2',3-difucosyllactose (diFL), lactose-N-fucopentaose I (LNFP I), Gal-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (Gal-LNFP I), GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc (GalNAc-LNFP I), lactose-N-neofucopentaose I (LNnFP I), lactose-N-fucopentaose II (LNFP II), lactose-N-fucopentaose III (LNFP III), lactose-N-fucopentaose V (LNFP V), lactose-N-fucopentaose VI (LNFP VI), lactose-N-neofucopentaose V, lactose-N-difucohexaose I (LNDFH-I), lactose-N-difucohexaose II (LNDFH-II), Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,2-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Fuc-a1,4-(Fuc-a1,2-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, monofucosyllactose-N-hexaose-III, difucosyllactose-N-hexaose (a), difucosyllactose-N-hexaose, difucosyllactose-N-neohexaose, trifucosyllactose-N-hexaose, a1,3- Galactosyl-3-fucosyllactose, Gal-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, GalNAc-a1,3-(Fuc-a1,2-)Gal-b1,4-(Fuc-a1,3-)Glc, 2-fucosyllactulose, 3-fucosyl-N-acetyllactosamine, 2'-fucosyl-N-acetyllactosamine, difucosyl-N-acetyllactosamine, 4-fucosyllactose-N-disaccharide, 2'-fucosyllactose-N-disaccharide, difucosyllactose-N-disaccharide and GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc; sialylated oligosaccharides, selected from 3'-sialyllactose (3'SL), 6'-sialyllactose (6'SL), 8'-sialyllactose (8'SL), 3,6-disialyllactose (Neu5Ac-α2,3-(Neu5Ac-α2,6)-Gal-β1,4-Glc), 6,6'-disialyllactose (Neu5Ac-α2,6-Gal-β1,4-(Neu5Ac-α2,6)-Glc), 8,3-disialyllactose (Neu5Ac-α2,8-Neu5Ac-α2,3-Gal-β1,4-Glc), 6'-sialyllactosamine, 3'-sialyllactosamine, sialyllactose-N-tetraose a (LSTa), sialyllactose-N-tetraose b (LSTb) , sialyllactose-N-tetraose c (LSTc), sialyllactose-N-tetraose d (LSTd), disialyllactose-N-tetraose, disialyllactose-N-neotetraose, monosialyllactose-N-hexaose, disialyllactose-N-hexaose I, disialyllactose-N-hexaose II, monosialyllactose-N-neohexaose I, monosialyllactose-N-neohexaose II, disialyllactose-N-neohexaose, 3'-sialyl-3-fucosyllactose, fucodisialyllactose-N-hexaose, disialylmonofucosyllactose-N-neohexaose, sialyllactose-N-fucohexaose II, disialyllactose-N-fucopentaose II, monofucosyldisialyllactose-N-tetraose, Neu5Ac-a2,3-Gal-b1,4-GlcNAc-b1,3-Gal, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-KDO-lactose, 3'-KDO-lactosamine, 3'-KDO-6'-sialyllactose, 3'KDO-8-sialyllactose, KDO-2,3Galβ-1,3GalNacβ-1,3Galα-1,4Galβ-1,4Gal, KDO-2,3Galβ-1,3GlcNacβ-1,3Galβ-1,4Glc, KDO-2,3Galβ-1,4GlcNacβ-1,3Galβ-1,4Glc, 3'-KDO-3-fucose Neu5Ac-a2,8-Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal, 3'-sialyl-2'-fucosyllactose, 6'-sialyl-2'-fucosyllactose, 6'-sialyl-3-fucosyllactose, 3'-sialyl-3-fucosyllactose, Neu5Ac-a2,6-(Neu5Ac-a2,3-)Gal-b1,4-Glc, 3'-sialyl-3-fucosyllactosamine, Fuc-a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc, 6'-sialyl-N-disaccharide, 3'-sialyl-N-disaccharide, N eu5Ac-a2,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,3-Gal-b1,4-(Fuc-a 1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,4-(Fuc-a1,3-)GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,4-G lcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1, 4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-(Fuc-a1,3-)Glc, Neu5Ac-a2,3-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc,Fuc -a1,4-(Neu5Ac-a2,3-Gal-b1,3-)GlcNAc-b1,3-Gal-b1,4-Glc, Neu5Ac-a2,6-(Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-(Fuc-a1,3-)Glc , Neu5Ac-a2,6-(Neu5Ac-a2,6-(Fuc-a1,2-)Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc, Neu5Ac-a2,6-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, and any combination thereof; neutral (uncharged) oligosaccharides containing N-acetylglucosamine selected from lacto-N-triose II (LN3), lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), 6'-galactosyl Lactose, 4'-galactosyl lactose, 3'-galactosyl lactose, GlcNAc-b1,6-Gal-b1,4-Glc, lactose-N-hexaose (LNH), lactose-N-neohexaose (LNnH), p-lactose-N-hexaose (pLNH), p-lactose-N-neohexaose (pLNnH), GlcNAc-b1,6-(GlcNAc-b1,3-)Gal-b1,4-Glc, lactose-N-pentaose (LN5), lactose-N-neopentaose, p-lactose-N-pentaose, p-lactose-N-neopentaose, lactose-N-neopentaose I, lactose-N-heptaose (LN7), lactose-N-neoheptaose, p-lactose-N-neoheptaose, p-lactose-N-heptaose, lactose -N-octaose (LNO), lactose-N-neooctaose, allolactose-N-octaose, p-lactose-N-octaose, allolactose-N-neooctaose, novel lactose-N-neooctaose, p-lactose-N-neooctaose (pLNnO), allolactose-N-nonaose, novel lactose-N-nonaose, lactose-N-nonaose (LN9), lactose-N-decaose, allolactose-N-decaose, novel lactose-N-decaose, lactose-N-neodecaose, p-lactose-N-neodecaose (pLNnD), a1,3-galactosyl lactose-N-neotetraose, GlcNAc-b1,3-Gal-b1,3-GlcNAc-b1,3-Gal-b1,4-Glc, GlcNAc-b1,6-(Gal-b1,4-GlcNAc-b1,3-)Gal-b1,4-Glc and GlcNAc-b1,6-(Gal-b1,3-GlcNAc-b1,3-)Gal-b1,4-Glc; mammalian milk oligosaccharide (MMO); human milk oligosaccharide (HMO); animal oligosaccharide selected from N-glycans and O-glycans; plant oligosaccharide selected from N-glycans and O-glycans.

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