Belt reactor manufacturing process for oligomer synthesis

Abstract

A method and apparatus for synthesis of oligomers such as oligonucleotides and peptides are disclosed. The method employs a device including at least one deprotection station to carry out a step of deprotection, at least one coupling station to carry out a step of coupling, optionally, one or more oxidation and / or thiolation stations to carry out a step of oxidation or thiolation, optionally, one or more capping stations to carry out a step of capping, and, optionally, one or more washing stations to carry out a step of washing. A plurality of solid phase material for oligomer synthesis is moved to the stations via a conveyor device, wherein a fluid delivery device acts upon said solid phase material to produce an oligomer.

Description

BELT REACTOR MANUFACTURING PROCESS FOR OLIGOMER SYNTHESIS

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 482,675, filed on February 1, 2023, the contents of which are incorporated by reference in their entirety.Field of the Disclosure

[0002] The present disclosure relates to an apparatus and a method for solid phase synthesis of oligomers such as oligonucleotides and peptides.Background of the Disclosure

[0003] Oligonucleotides and peptides are biological polymers that play pivotal roles in the biological, medical, and pharmaceutical industries (Wan et al., J. Med. Chem. 2016, 59, 21, 9645-9667). As such, major efforts have been dedicated to improving their efficient synthesis, particularly through solid phase methodologies (Merrifield et al., “Solid Phase Synthesis” Science 1986, 232, 341-347; Itakura et al., “Synthesis and Use of Synthetic Oligonucleotides” Annual Review of Biochemistry 1984, 53(1), 323-356).

[0004] Common methods for producing oligonucleotides include phosphoramidite, phosphortri ester, or H-phosphonate methods (Verma et al., “Modified Oligonucleotides: Synthesis and Strategy' for Users” Annu. Rev. Biochem. 1998, 67, 99-134). In these cases, the activated phosphorous atom of one nucleotide reacts with the 5 '-hydroxy group of the sugar in another nucleotide to form a new covalent bond, effectively linking the nucleotides together. Oxidation of the newly formed phosphite to a phosphate further stabilizes the connection. After deprotection of the 5’-hydroxy group of the newly introduced nucleotide, these steps can be repeated, thereby growing the nucleotide chain with each step, until the desired sequence is assembled. To increase efficiency and throughput, oftentimes a solid phase synthesis methodology’ can be employed.

[0005] Akin to the synthesis of oligonucleotides, peptides can be produced through similar strategies (Jaradat, “Thirteen decades of peptide synthesis: key developments in solid phase peptide synthesis and amide bond formation utilized in peptide ligation” Amino Acids 2018, 50, 39-68). The C-terminus of an N-protected amino acid can be activated through appropriate reagents known in the art and coupled to the N-terminus of another amino acid, thus forming anew amide bond. After deprotection of the N-terminus of the newly added amino acid, the process can be repeated with the same or a different amino acid to grow the peptide chain until the desired sequence is prepared.

[0006] Solid phase synthesis ty pically involves the immobilization of organic molecules through covalent bonds on solid phase material and their subsequent chemical manipulation while the connection to the solid support is maintained (Dorwald, “Organic Synthesis on Solid Phase: Supports, Linkers, Reactions” Wiley-VCH: Weinheim, 2002). Traditional apparatuses for the synthesis of oligomers such as oligonucleotides or peptides (z.e., solid phase synthesizers) often employ this strategy and adopt a flow-through design. A column is packed with solid phase material, and a mobile phase containing the requisite reactant or reagent is passed through. As the mobile phase passes through the column, the chemical agent affects the desired manipulations on the organic molecules. Albeit practical, there are disadvantages of such setups such as high cost and difficulty' of upscaling the process.

[0007] As the amount of oligonucleotide or peptide produced correlates with the amount of solid phase material in the column, one way to increase production volume is to expand the column size. However, due to physical constraints relating to flow distribution, operating back pressure, and pore diffusion, the maximum column size is limited (Moore et al. , “Overcoming Backpressure Problems during Solid-Phase Synthesis of Oligonucleotides” Org. Proc. Res. Dev. 2004, 8(2). 271-274). Therefore, an increase in production volume is often achieved through either manufacturing several batches sequentially7or the use of multiple columns, conducting all steps in parallel. The latter results in an increased amount of pipes, pumps, valves, and other materials required, which adds to the production cost and consequently renders the scale-up challenging.

[0008] Thus, a need remains for an improved workflow' for a more streamlined, cost- effective, and scalable synthesis of oligonucleotides and peptides. The present disclosure provides, inter alia, a continuous manufacturing approach to overcome the limitations of batchwise production and to facilitate the production of biopolymers on larger scales.Summary of The Disclosure

[0009] The present disclosure provides, inter alia, an improved solid phase synthesizer for continuous or quasi-continuous manufacture of oligomers, as well as methods and systems for continuous or quasi-continuous manufacture of oligomers such as oligonucleotides and peptides.

[0010] In some embodiments, an apparatus for the synthesis of oligomers is provided. In some embodiments, the apparatus comprises at least one deprotection station to carry' out a step of deprotection, at least one coupling station to carry7out a step of coupling, optionally one or more oxidation and / or thiolation stations to carry out a step of oxidation or thiolation,optionally, one or more capping stations to carry out a step of capping, and, optionally, one or more washing stations to carry out a step of washing. In some embodiments embodiments, one or more solid phase materials are moved to each station via a conveyor device. At each station, a fluid delivery device dispenses a reagent, a reactant, or a washing fluid to the solid phase material to produce an oligomer (e.g., an oligonucleotide or peptide).

[0011] In some embodiments, an apparatus for the synthesis of oligonucleotides is provided. The apparatus comprises at least one deprotection station to carry out a step of deprotection, at least one coupling station to carry out a step of coupling, at least one oxidation and / or thiolation station to cany' out a step of oxidation or thiolation, optionally, one or more capping stations to carry out a step of capping, and, optionally, one or more washing stations to carry out a step of washing. One or more solid phase materials are moved to each station via a conveyor device. At each station, a fluid delivery device dispenses a reagent, a reactant, or a washing fluid to the solid phase material to produce an oligonucleotide.

[0012] In some embodiments, a method of continuous or quasi-continuous oligonucleotide synthesis is provided, comprising (a) providing a solid phase material and an apparatus for oligonucleotide synthesis, said apparatus comprising at least one deprotection station to carry out a step of deprotection; at least one coupling station to carry out a step of coupling; at least one oxidation and / or thiolation station to cany' out a step of oxidation or thiolation; optionally, one or more capping stations to cany out a step of capping, and, optionally, one or more washing stations to carry out a step of washing; wherein the apparatus is configured to act on one or more solid phase materials at each station to produce an oligonucleotide; and wherein each station is at a fixed position in the apparatus and configured to deliver a reagent, a reactant, or a washing fluid to a moving solid phase material via a fluid delivery device, and (b) initiating movement of the solid phase material through the stations of the apparatus.

[0013] In some embodiments, an apparatus for the synthesis of peptides is provided. The apparatus comprises at least one deprotection station to cany' out a step of deprotection, at least one coupling station to carry out a step of coupling, optionally one or more capping stations to carry out a step of capping, and, optionally, one or more washing stations to carry out a step of washing. One or more solid phase materials are moved to each station via a conveyor device. At each station, a fluid delivery device delivers a reagent, a reactant, or a washing fluid to the solid phase material to produce a peptide.

[0014] In some embodiments, a method of continuous or quasi-continuous peptide synthesis is provided, comprising (a) providing a solid phase material and an apparatus for peptidesynthesis, said apparatus comprising at least one deprotection station to carry out a step of deprotection; at least one coupling station to carry out a step of coupling; optionally one or more capping stations to carry out a step of capping, and optionally, one or more washing stations to carry out a step of washing; wherein the apparatus is configured to act on one or more solid phase materials at each station to produce a peptide; and wherein each station is at a fixed position in the apparatus and configured to deliver a reagent, a reactant, or a washing fluid to a moving solid phase material via a fluid delivery device, and (b) initiating movement of the solid phase material through the stations of the apparatus.

[0015] In some embodiments, a solid phase belt is used as the solid phase material and moved to each station via a conveyor device. In some embodiments, the solid phase belt is comprised of linked cassettes. In some embodiments, the solid phase belt is comprised of linked cassettes forming a continuous solid phase belt. In some embodiments, the cassettes are detachable. In some embodiments, a flexible section of the solid phase belt or the cassette is flat, folded, layered, rolled, arranged in a spiral, or otherwise configured to increase the surface area. In some embodiments, the solid phase belt comprises one or more of polymers, glass, carbon, graphene, and graphene oxide. In some embodiments, the solid phase belt comprises a nonwoven material. In some embodiments, the solid phase material is treated to increase or add binding capabilities to one or more oligomer.Brief Description of the Drawings

[0016] The following is a brief description of the drawings, presented for the purpose of illustrating the embodiments disclosed herein and not meant to impose a limit on the present disclosure.

[0017] Fig. 1 shows an exemplary process to synthesize oligonucleotides with solid phase particles using a belt reactor that includes multiple modules.

[0018] Fig. 2 shows an example of a single module of a belt reactor operating with solid phase particles and comprising several horizontally arranged stations.

[0019] Fig. 3 show s an example of a single station within a module of a belt reactor.

[0020] Fig. 4 shows an example of a single module of a belt reactor, comprising several stations that are arranged around a moving rotary cylinder. The solid phase material could be particles or a nonwovens.

[0021] Fig. 5 shows an exemplary process to synthesize oligonucleotides with a solid phase belt that includes multiple modules.

[0022] Fig. 6 shows an example of a single module of a belt reactor that uses a solid phase belt.

[0023] Fig. 7 shows an example of a single station within a module of a belt reactor operating with a solid phase belt.

[0024] Fig. 8 shows an example of a single module of a belt reactor operating with cassettes and having several horizontally arranged stations.

[0025] Fig. 9 show s six examples of cassettes: (a) a single sheet of a flexible band arranged a cassette; (b) several sheets of a flexible band of the same material arranged in a cassette; (c) several sheets of flexible bands of tw o different materials arranged in a cassette; (d) a single sheet of a solid phase belt arranged in a folded manner in a cassette; (e) several sheets of a solid phase belt of the same material arranged in a folded manner inside a cassette; and (f) several sheets of two solid phase belts of different materials arranged in a folded manner inside a cassette.Detailed Description

[0026] To better understand the details of the disclosure, certain terms are first defined herein. Also, the contents of all references, patents, and published patent applications cited throughout this application, as well as the figures, are incorporated herein by reference in their entirety for all purposes. To the extent a reference statement contradicts a statement in the instant disclosure, the instant disclosure will control.Definitions

[0027] As used herein, the singular terms “a,’' “an,’‘ and “the” include the plural reference unless the context clearly indicates otherwise. The phrase “and / or,” as used herein, means “either or both” of the elements so conjoined, i.e.. elements that are conjunctively present in some cases and disjunctively present in other cases. Thus, as a non-limiting example, “A and / or B”, when used in conjunction with open-ended language such as “comprising” can refer, in some embodiments, to A only (optionally including elements other than B); in other embodiments, to B only (optionally including elements other than A); in yet other embodiments, to both A and B (optionally including other elements); etc.

[0028] As used herein, “at least one” means one or more of the elements in the list of elements, but not necessarily including at least one of each and every' element specifically listed w ithin the list of elements and not excluding any combinations of elements in the list of elements.

[0029] As used herein, “about” when used in connection with amounts or ratios, include the value of a specified amount or ratio that is recognized by one of ordinary skill in the art to provide a desired function equivalent to that obtained from the specified amount or ratio. The term “about” may refer to an acceptable error for a particular value as determined by one of skill in the art, which depends in part on how the values is measured or determined. In some embodiments, the term “about” means within 5%. 4%, 3%, 2%, 1%, or 0.5% of a given value or range.

[0030] As used herein, the term “agent” encompasses any chemical element or compound. Examples of agents include but are not limited to reactants, reagents, and washing fluids.

[0031] As used herein, the term “amino acid” refers to a molecule containing both an amine group and a carboxyl group. Suitable amino acids include, without limitation, both the D- and L-isomers of the naturally occurring amino acids, as well as non-naturally occurring amino acids prepared by organic synthesis or other metabolic routes. In some embodiments, a single “amino acid” might have multiple sidechain moieties, as available per an extended aliphatic or aromatic backbone scaffold. In some embodiments, one or more functional groups of the amino acid may be protected with appropriate protecting groups.

[0032] As used herein, the term “belt reactor” refers to an apparatus that provides for the continuous manufacture of solid phase supported oligomers, e.g., biological polymers, peptides, and oligonucleotides, moving along a conveyance system. A belt reactor can comprise one or more modules that act on solid phase material (e.g., a solid phase belt), which may be directly deposited on a conveyor device without the need for a reaction vessel or which may be provided in cassettes, vessels or other surfaces affixed to the conveyor device.

[0033] As used herein, the term “module” refers to a part of a belt reactor. A module can comprise one or more stations, wherein each module is configured to let the oligomer grow by one monomer or preexisting building block thereof. Examples of monomers include nucleotides in the case of oligonucleotide synthesis and amino acids in the case of polypeptide synthesis. Examples of preexisting building blocks include oligonucleotides in the case of oligonucleotide synthesis and peptides in the case of peptide synthesis. Modules can be arranged in sequence to synthesize a specific oligomer sequence. Modules may exhibit different lengths and may be spaced at different distances from each other, depending on the target reaction and exposure time before the solid phase material arrives at the next module. A single module can be used more than once (e.g., multiple copies of the same type of module may be present in the belt reactor). Alternatively, the solid phase material may beacted upon by one and the same module more than once by feeding the solid phase material back to the module after the first or any subsequent pass.

[0034] As used herein, the term '‘cassette’’ refers to a segment of a solid phase belt, wherein cassettes are jointly liked to other cassettes or spacers to form a continuous chain. A cassette may be independently detachable from the remainder of the solid phase belt at any point of the solid phase belt. A single cassette may comprise a flexible band and a rigid frame that provides stability. The flexible band of the cassette may comprise a woven or nonwoven material, which exhibits functional groups on its surface that allow for solid phase synthesis of oligomers. Exemplary' flexible band materials of the cassette include but are not limited to polymers, glass, carbon, graphene, and graphene oxide. The flexible band of the cassette may comprise a single sheet, multiple sheets of the same material, and / or multiple sheets of different materials. The configuration of the flexible band of the cassette may be flat, folded, layered, rolled, arranged in a spiral, or otherwise configured to increase the surface area of the flexible band. The rigid section of the cassette may comprise materials such as metal, polymer, wood, glass, and other materials that provide mechanic stability. The rigid section of the cassette may be configured to retain the shape of the flexible band it compnses. The rigid section of the cassette may be made of materials that are inert to the reagents and reactants to which the cassette is exposed during operation of the belt reactor. Cassettes may further be configured to be sealable to the outside, thus creating a barrier between the flexible section of the cassette and the ambient atmosphere.

[0035] The term '‘catalyst” refers to a chemical compound that accelerates the rate at which a chemical reaction occurs. A catalyst is not turned into a product itself or otherwise changed or consumed at the completion of the chemical reaction. After a catalyst participates in a chemical reaction, because it is unchanged, it may participate in further chemical reactions, acting on additional reagents or reactants to create additional products.

[0036] As used herein, the term “conveyor device” refers to a part of a belt reactor that transports a solid phase material from one station to another. The conveyor device may also be used to transport the solid phase material from one module to another. Example of such a conveyor device include but are not limited to a conveyor belt and conveyor rollers. The device may be configured for continuous movement of a belt reactor and / or for periodic slowing or pausing of the belt reactor.

[0037] As used herein, the term “modified nucleotide,” “chemically modified nucleotide”, or “chemically modified variant” is consistent with how that term is defined in the art and can include, but is not limited to, modified variants of DNA and RNA such as 2'-modifiednucleotides, inverted nucleotides, modified nucleobase-comprising nucleotides, bridged nucleotides, peptide nucleic acid monomers (e.g, repeating N-(2-aminoethyl)glycine (AEG) groups) that form peptide nucleic acids (PNAs), unlocked nucleotides, 2',3'seco nucleotide mimics (unlocked nucleobase analogues), locked nucleotides, 3'-O-methoxy (2' intemucleoside linked) nucleotides, 2'-F-Arabino nucleotides, 5'-Me, 2'-fluoro nucleotides, morpholino nucleotides, abasic nucleotides (also referred to as an “abasic site”), vinyl phosphonate-containing nucleotides, and cyclopropyl phosphonate-containing nucleotides. Commonly used modified nucleotides include 2'-modified nucleotides (z.e., a nucleotide with a group other than -H or -OH at the 2' position of the five-membered sugar ring) which include, but are not limited to, 2'-O-methyl nucleotides (also referred to as 2'-methoxy nucleotides). 2'-fluoro nucleotides (also referred to as 2'-deoxy-2'-fluoro nucleotides), 2'- methoxy ethyl (2'-O-2-methoxylethyl) nucleotides (also referred to as 2'-M0E nucleotides), 2'-amino nucleotides, and 2'-alkyl nucleotides. In some embodiments, a modified nucleotide comprises a modified nucleobase such as 5-substituted pyrimidines, 6-azapyrimidines and N- 2, N-6 and 0-6 substituted purines, (e.g, 2aminopropyladenine, 5-propynyluracil, or 5- propynylcytosine). 5-methylcytosine (5-me-C). 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-alkyl (e.g, 6-methyl, 6-ethyL 6-isopropyl, or 6-n-butyl) derivatives of adenine and guanine, 2-alkyl (e.g., 2-methyl, 2-ethyl, 2-isopropyl, or 2-n-butyl) and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine, 2- thiocytosine. 5-halouracil, cytosine, 5-propynyl uracil, 5-propynyl cytosine, 6-azo uracil, 6- azo cytosine, 6-azo thymine, 5-uracil (pseudouracil), 4thiouracil, 8-halo, 8amino, 8- sulfhydryl, 8-thioalkyl. 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo (e.g., 5-bromo), 5 -trifluoromethyl, and other 5-substituted uracils and cytosines, 7- methylguanine and 7-methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine. 7— deazaadenine, 3-deazaguanine, and 3-deazaadenine. In some embodiments, an oligonucleotide may comprise modified intemucleoside linkages or backbones. Such modified linkages include, but are not limited to, phosphorothioates, chiral phosphorothioates. thiophosphates, phosphorodithioates, phosphotriesters, aminoalkylphosphotriesters. alkyl phosphonates (e.g, methyl phosphonates or 3'-alkylene phosphonates), chiral phosphonates, phosphinates, phosphoramidates (e.g, 3'-amino phosphoramidate, aminoalkylphosphoramidates, or thionophosphoramidates), thionoalkyl- phosphonates, thionoalkylphosphotriesters, morpholino linkages, boranophosphates having normal 3'-5' linkages, 2'-5' linked analogs of boranophosphates, or boranophosphates having inverted polarity wherein the adjacent pairs of nucleoside units are linked 3'-5' to 5'-3' or 2'-5'to 5 '-2'. In some embodiments, a modified intemucleoside linkage or backbone lacks a phosphorus atom. Modified intemucleoside linkages lacking a phosphorus atom include, but are not limited to, short chain alkyl or cycloalkyd inter-sugar linkages, mixed heteroatom and alkyl or cycloalkyl inter-sugar linkages, or one or more short chain heteroatomic or heterocyclic inter-sugar linkages. In some embodiments, modified intemucleoside backbones include, but are not limited to, siloxane backbones, sulfide backbones, sulfoxide backbones, sulfone backbones, formacetyl and thioformacetyl backbones, methylene formacetyl and thioformacetyl backbones, alkene-containing backbones, sulfamate backbones, methyleneimino and methylenehydrazino backbones, sulfonate and sulfonamide backbones, amide backbones, and other backbones having mixed N, O, S, and CH2 components. As is known and reported in the art, multiple modifications may be applied to a single nucleotide, and an oligonucleotide may feature various different types of nucleotides (including natural nucleotides, modified nucleotides, and combinations thereof).

[0038] As used herein, the term “monomer” refers to a single biological building block such as a single nucleotide or a single amino acid. “Monomer” may be used in the context of an existing oligomer sequence and refer to a single building block therein or to a new building block such as a nucleotide or an amino acid that is to be added to a solid-bound monomer or solid-bound oligomer.

[0039] As used herein, the term “nucleotide” includes traditional natural nucleotides as well as chemically modified variants thereof. A nucleotide can be synthetically manufactured.

[0040] As used herein, the term “oligomer” refers to a molecule that comprises at least two monomers and is meant to encompass biological polymers such as those comprising an oligonucleotide or a peptide.

[0041] As used herein, the term “oligonucleotide” refers to a compound, comprising natural nucleotides such as DNA and RNA or chemically modified variants thereof. Examples include, but are not limited to DNA, RNA, mRNA (messenger RNA), hybrids of DNA and / or RNA (this includes chemically modified variants thereof), RNAi (RNA and / or chemically modified RNA that can induce an RNA interference mechanism), siRNA (small interfering RNA), shRNA (short hairpin-RNA). miRNA (micro-RNA), antisense RNA, ribozymes, catalytic DNA, RNA that induce triple helix formation, aptamers, and vectors.

[0042] As used herein, the term “peptide” refers to a compound, wherein two or more amino acids are covalently attached together. The term “peptide” is meant to be used interchangeably with the terms “protein”, “oligopeptide”, and “polypeptide”. Peptides may be made up of naturally occurring and / or synthetic amino acids.

[0043] As used, herein, the term “quasi-continuous” refers to a sequence of operations during which the flow of product material (e.g., solid phase material or liquid feeds) takes place, in principle, continuously during certain periods but wherein the flow can be interrupted for one or more period of time. For instance, when operation of the belt reactor comprises the use of cassettes, the process as a whole is continuous but might involve interruption, e.g., when moving between cassettes or to exclude cassettes.

[0044] As used herein, the term “reactant” refers to agents that comprise a monomer or preexisting building block in an activated form, a protected form, or a form that allows the monomer or preexisting building block to be coupled to another monomer or oligomer through a chemical reaction. In the case of oligonucleotide synthesis, examples include but are not limited to nucleoside phosphoramidites, phosphortriesters, and H-phosphonates. In the case of peptide synthesis, examples include but are not limited to amino acids, partially protected amino acids, N-protected amino acids, and C-protected amino acids.

[0045] As used herein, the term “reagent’’ refers to chemical compounds that are used to assist in, facilitate, or perform a chemical reaction. Examples include but are not limited to activators, catalysts, oxidants, thiolation agents, and capping agents.

[0046] As used herein, the term “solid phase belt” refers to a type of solid phase material comprising a flexible continuous band made from a permeable material that exhibits functional groups on its surface that allow for solid phase synthesis of oligomers. Examples of applicable materials for the solid phase belt include but are not limited to polymers, glass, carbon, graphene, and graphene oxide. A solid phase belt may comprise either a woven or a nonwoven fabric. A solid phase belt may also be comprised of interconnecting cassettes that are joined to form a continuous chain. In some embodiments, the solid phase belt comprises one or more interconnected cassettes. In some embodiments, the solid phase belt comprises ten or more interconnected cassettes. In some embodiments, the solid phase belt comprises fifty or more interconnected cassettes.

[0047] As used herein, the term “solid phase material” refers to polymers, resins, controlled porous glass, graphene, graphene oxide, glass, and other materials that can be used as solid supports for the solid phase synthesis of oligomers such as oligonucleotides and peptides. Examples of common solid phase materials include but are not limited to Nittophase®, Tentagel®, and a controlled porous glass (CPG).

[0048] As used herein, the term “station” refers to a distinct segment of a module, where a specific step is carried out. Examples of such steps include but are not limited to deprotection, coupling, oxidation, thiolation, capping, and washing. A single module cancontain one or more stations of the same type. Stations may vary with respect to length, construction, and other variables to accommodate the specific requirements of the step that is carried out at the station.AbbreviationsA AdenineAloe AllyloxycarbonylATR Attenuated total reflectanceBn BenzylBOM BenzyloxymethylBOP Benzotriazol- 1 -yloxytris(dimethylamino)phosphonium hexafluorophosphateC CytosineCbz Carbobenzyloxy cGMP Current Good Manufacturing PracticeCOMU (l-Cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino- morpholino-carbenium hexafluorophosphateCPG Controlled porous glassCSA Camphorsulfonic acidDCC N,N'-dicyclohexylcarbodiimideDDTT 3-(Dimethylaminomethylidene)amino-3H-l,2,4-dithiazole-3-thioneDIC DiisopropylcarbodiimideDMB 2,4-DimethoxybenzylDMF N,N- Di methyl formamideDMT 4,4'-DimethoxytritylDNA Deoxyribonucleic acidECTFE EthylenechlorotrifluoroethyleneEDC 1 -Ethyl-3-(3-dimethylaminopropyl)carbodiimideEPDM Ethylene propylene diene monomer rubberFmoc 9-Fluorenylmethoxy carbonylG GuanineHATU l-[Bis(dimethylamino)methylene]-lH-l,2,3-triazolo[4,5- b]pyridinium 3-oxide hexafluorophosphateHBTU O-(Benzotriazol- 1 -yl)-N,N,N’ ,N'-tetramethy luronium hexafluorophosphateHO At l-Hydroxy-7-azabenzotriazoleHOBt 1 -HydroxybenzotriazoleIR InfraredNMI 1 -MethylimidazoleNMR Nuclear magnetic resonanceLCMS Liquid chromatography mass spectrometryPA PolyamidePE PolyethylenePEEK Polyether ether ketonePET Polyethylene terephthalatePP PolypropylenePTFE PolytetrafluoroethylenePTS A 4-Methylbenzene-l -sulfonic acidPU PolyurethanePVC Polyvinyl chloridePVDF Polyvinylidene fluoridePyBOP Benzotriazol- 1 -yloxy tripyrrolidinophosphonium hexafluorophosphateRMV Rubber-modified vinylRNA Ribonucleic acidSBR Styrene-butadiene rubberSEM 2-TrimethylsilylethoxymethylT ThymineT3P Propylphosphonic anhydrideTATU O-(7 - Azabenzotriazol- 1 -yl)- N,N,N’ ,N’ -tetramethy luronium tetrafluoroborateTBDPS / e / 7-ButyldiphenylsilylTBS / e / 7-ButyldimethylsilylTBTU O-(Benzotriazol- 1 -yl)- N,N,N’ ,N’ -tetramethyluronium tetrafluoroborateTPE Thermoplastic elastomerU UracilManufacturing of Oligomers using a Belt Reactor

[0049] In some embodiments, the disclosure provides a method and apparatus for continuous or quasi-continuous manufacture of oligomers such as oligonucleotides and peptides. The steps of the manufacturing process (which can be carried out by the apparatus) comprise at least one deprotection station to carry out a step of deprotection, at least one coupling station to carry out a step of coupling, optionally, one or more oxidation / thiolation stations to carryout a step of oxidation or thiolation, optionally, one or more capping stations to carry out a step of capping, and optionally, one or more washing stations to carry' out a step of washing. One or more solid phase materials are moved between the stations via a conveyor device, and, at each station, a fluid delivery' device provides a reactant or reagent to the solid phase material.

[0050] In some embodiments, the belt reactor comprises one or more modules. The number of modules and the configuration thereof can be adjusted based on the desired oligomer, production volume, manufacturing speed, and / or other production parameters and requirements. In some embodiments, production occurs in a single module (e.g., with all stations for the preparation of the entire oligomer of interest present in the module). In some embodiments, production occurs in more than one module (e.g., with each module providing the steps for a portion of the process, e.g, the steps for adding a single amino acid or nucleotide to an oligomer).

[0051] In some embodiments, the synthesis of an oligomer is achieved through a linear arrangement of modules, whereby each module adds a nucleotide or ammo acid to the growing chain. In some embodiments, the synthesis of an oligomer is achieved through a linear arrangement of modules, whereby each module adds a nucleotide, amino acid, or preexisting building block to the growing chain.

[0052] In some embodiments, the synthesis of an oligomer is achieved through a non-linear arrangement of modules, wherein each module adds a monomer or preexisting building block to the growing chain, and wherein one or more modules are used more than once. In some embodiments, the synthesis of an oligonucleotide or peptide is achieved through a non-linear arrangement of modules, wherein each module adds a nucleotide, amino acid, or preexisting building block to the growing chain, and wherein one or more modules are used more than once. In some embodiments, the solid phase material undergoes one or more modules more than once, wherein the solid phase material is resupplied to an earlier module at least once toachieve the desired sequence. In some embodiments, multiple copies of a desired oligomer are produced in parallel in the same or separate modules.

[0053] In some embodiments more than one solid phase belt is acted upon in a module. In some embodiments, two or more solid phase belts are vertically stacked on top of each other within one module. In some embodiments, two or more solid phase belts are horizontally stacked next to each other within one module. In some embodiments, one or more fluid delivery device dispenses reactant, reagent, or washing fluid to two or more stacked solid phase belts simultaneously. In some embodiments, one or more fluid delivery device dispenses reactant, reagent, or washing fluid to two or more stacked solid phase belts sequentially, wherein the reactant, reagent, or washing fluid permeates through one belt first before permeating through the other belt(s). In some embodiments, reactant, reagent, or washing is dispenses at a first solid phase belt, recovered, and the dispensed at an additional solid phase belt within the same module.

[0054] In some embodiments, a module comprises one or more stations that are arranged sequentially in a horizontal manner. In some embodiments, a module comprises one or more stations that are arranged sequentially in a stacked or vertical manner. In some embodiments, a module comprises one or more stations that are arranged sequentially around a moving rotary7cylinder.

[0055] In some embodiments, a buffer tank is used in between two modules. In this case, the solid phase material from the end of one module is collected in the buffer tank and supplied to the start of the next module. In some embodiments, the buffer tank also acts as a reaction vessel to carry7out a specific step. In some embodiments, the buffer tank carries out a deprotection step. In some embodiments, a separate reaction vessel is used to carry out a specific step of the sequence. In some embodiments, a separate reaction vessel is used to carry out the deprotection step. In some embodiments, the buffer tank is used in a belt reactor with solid phase particles.

[0056] In some embodiments, no buffer tank is used, and the solid phase material is directly- transferred from the end of one module to the start of the next module.

[0057] In some embodiments, the number, types, and arrangement of stations in a module can differ depending on the desired oligomer sequence, desired level of purity, projected cost, and other parameters. In some embodiments, there is more than one station carry ing out the same step within the same module. In some embodiments, one or more washing steps are conducted after deprotection, coupling, oxidation, thiolation, and / or capping steps. In other embodiments, some steps are conducted in sequence without a washing step in between. Insome embodiments, each module of a belt reactor comprises a capping step to prevent the production of oligomer sequences with missing nucleotides or amino acids. In other embodiments, not every module comprises a capping step. In some embodiments, each module of a belt reactor comprises an oxidation or thiolation step. In other embodiments, not every module comprises an oxidation step or thiolation step.

[0058] In some embodiments, the length of a station can be varied to accommodate differences in reaction kinetics of the deprotection, coupling, oxidation, thiolation, capping, and / or washing step. Increasing the length of a station can lead to an increased time the solid phase material on the conveyor device or the cassette takes to reach the next station. Consequently, the solid phase material may be exposed to the reactant, reagent, or washing fluid dispensed at that station for a longer period of time. In some embodiments, operation of the belt reactor comprises quasi-continuous movement of the conveyor device to allow for longer reaction time of one or more reactions at one or more stations. Increasing the length of stations where slow reactions are carried out may thus facilitate greater reaction completion and / or increase the purity of the synthesized oligomer. In some embodiments, increasing the length of a station and increasing the speed at which the conveyor belt transports the solid phase material can lead to an increased amount of material throughput. In some embodiments, increasing the height and / or width of the solid phase material on the conveyor device may increase material throughput.

[0059] In some embodiments, the speed at which the conveyor device transports the solid phase material can be varied to accommodate differences in reaction kinetics of the deprotection, coupling, oxidation, thiolation, capping, and / or washing step. Decreasing the speed of the conveyor device can lead to an increased time the solid phase material on the conveyor device takes to reach the next station. Consequently, the solid phase material may be exposed to the reactant, reagent, or washing fluid dispensed at that station for a longer period of time. Decreasing the conveyor device speed at stations where slow reactions are carried out may thus facilitate greater reaction completion and / or increase the purity7of the synthesized oligomer. In some embodiments, increasing the speed at which the conveyor belt transports the solid phase material can lead to an increased amount of material throughput.

[0060] In some embodiments, each step is carried out in a distinct station where the requisite reagent, reactant, or washing fluid is dispensed onto the solid phase material on the conveyor device. The arrangement of stations within a module corresponds to the sequence of actions necessary7to affect the addition of a monomer to the solid-bound oligomer. To synthesize an oligomer, the solid phase material is acted upon sequentially by the appropriate modules thateach link one additional nucleotide or amino acid to the growing oligomer chain, respectively.

[0061] In some embodiments, the belt reactor is configured with a number of modules that corresponds to the total number of monomers in a desired oligomer. Each module may comprise all the stations needed for addition of a monomer (e.g., to add a nucleotide or an amino acid to a growing oligomer). For instance, to prepare an oligonucleotide of the sequence GAGA, four modules in total are employed and set up sequentially. Solid phase material is transported from one module to the next, in a linear succession, each adding one nucleotide or amino acid to the sequence. In some embodiments, different modules may be replaced in sequence to produce different oligomers. For instance, to switch from a sequence of GAGA to one of GGGA, different ”G ' and "A” producing modules may be aligned in the reactor.

[0062] In other embodiments, the belt reactor is configured with a number of modules that corresponds to the number of distinctly different monomers in a desired oligomer. For instance, to prepare an oligonucleotide of the sequence GAGA, two modules are employed in total. After the solid phase material has been acted upon by the first two modules, it is resupplied to the start of the first module to achieve the desired sequence.

[0063] In some embodiments, feeding tanks contain a reactant, reagent, or washing fluid. Reactants and reagents may be present in neat form or as a solution in an appropriate solvent. Feeding tanks may be connected to one or more fluid deli ven’ devices through coupling that provides for fluid communication. Examples include but are not limited to tubes, pipes, or functional equivalents thereof. If the same reagent, reactant, of washing fluid is used in more than one station, the belt reactor can be configured to allow one feeding tank to supply more than one station.

[0064] In some embodiments, the feeding tanks are refilled from supply wells. Reactants, reagents, and washing fluid in supply wells may be heated, cooled, shielded from light, stirred, or otherwise manipulated depending on the necessary storage conditions, stability , and specific properties of the respective agent.

[0065] In some embodiments, pumps are used to control the amount of reactant, reagent, or washing fluid dispensed. In some embodiments, the feeding tanks or supply wells are pressurized to dispense a reactant, reagent, or washing fluid upon opening of a valve. In some embodiments, one or more processors are programmed to control the operation of pumps that supply reactant, reagent, and washing fluid from supply wells and feeding tanks to the fluid delivery device.

[0066] In some embodiments, valves are used to control the flow of reactant, reagent, or washing fluid from the feeding tanks to one or more fluid delivery devices. The delivery of reactant, reagent, or washing fluid is controlled through the opening and closing of appropriate valves. In some embodiments, one or more processors are programmed to control the operation of valves that regulates delivery of reactant, reagent, and washing fluid from supply wells and feeding tanks to the fluid delivery device.

[0067] In some embodiments, one or more of reactant, reagent, and washing fluid are mixed prior to dispensing through the fluid delivery device. Premixing can be achieved through an additional premixing tank that is installed between the fluid delivery device and the supply well or the feeding tank. In some embodiments, one or more of reactant, reagent, and washing fluid are dispensed through the same fluid delivery device within a station, enabling mixing of one or more of reactant, reagent, and washing fluid upon release through the fluid delivery device.

[0068] In some embodiments, the fluid delivery device comprises a spray nozzle, a perforated nozzle, a perforated plate, or an open pipe for dispensing reagent, reactant, or washing fluid onto the solid phase material. Flow rate, outlet pressure, and spray angle of the fluid delivery device may be adjusted and controlled by process control. In some embodiments, the rates with which reactant, reagent, or washing fluid are dispensed onto the solid phase material is regulated through controlling the operation of pumps or valves. The spray angle of the fluid delivery device may be regulated through mechanical changes to the nozzle aperture and configured to produce the desired spray characteristic enabling optimal operation of a belt reactor with a specific conveyor width, conveyor speed, and solid phase material loading.

[0069] In some embodiments, a conveyor device transports the solid phase material. In some embodiments, the solid phase material, e.g., a solid phase belt, is integrated with the conveyor device. In some embodiments, the solid phase material, e.g., a solid phase belt, is disposed next to (e.g., on top of) the conveyor device. In some embodiments, the conveyor device that transports the solid phase material comprises rollers. In some embodiments, the conveyor device comprises a solid phase belt. In some embodiments, the conveyor device that transports the solid phase material is a conveyor belt. In some embodiments, the conveyor belt comprises a permeable material to allow liquid to pass through. In some embodiments, the conveyor belt serves as a filter. In some embodiments, the conveyor device comprises a filtration belt. In some embodiments, the conveyor device comprises a conveyor belt and a solid phase material (e.g., a solid phase belt). In some embodiments, the solid phase materialis attached to the conveyor belt. In some embodiments, the solid phase material is deposited on a conveyor belt (e.g. , to form a layer of solid phase material on top of a conveyer belt). In some embodiments, the conveyor belt comprises a single layer on the bottom of the solid phase material. In some embodiments, the conveyor belt comprises two layers that encompass the solid phase material on the top and the bottom. The material, width, length, thickness, tension, friction, porosity, and other characteristics of the conveyor belt may be selected based on, e.g.. the specific solid phase material, cassette, reactants, reagents, washing fluid, scale, ambient temperature, ambient pressure, and / or humidity at / with which the belt reactor is operated. In some embodiments, the conveyor belt adds mechanical stability to the solid phase material transported.

[0070] In some embodiments, the conveyor belt comprises a flat shape. In some embodiments, the conveyor belt comprises a folded (e.g., wave-like) structure. In some embodiments, the conveyor belt comprises an embossed surface. In some embodiments, the conveyor belt comprises a studded surface. In some embodiments, the conveyor belt comprises a diamond-shaped surface. In some embodiments, the conveyor belt comprises a sawtooth surface. In some embodiments, the conveyor belt comprises a grooved surface. In some embodiments, the conveyor belt comprises a cleated surface. In some embodiments, the conveyor belt comprises a curved surface. In some embodiments, the conveyor belt comprises a folded surface or a series of folded surfaces.

[0071] In some embodiments, the conveyor belt comprises a surface coating. In some embodiments, the conveyor belt comprises a PVC coating. In some embodiments, the conveyor belt comprises a rubber coating. In some embodiments, the conveyor belt comprises an aramid coating. In some embodiments, the conveyor belt comprises a polyester coating. In some embodiments, the conveyor belt comprises a natural rubber coating. In some embodiments, the conveyor belt comprises cellular foam coating. In some embodiments, the conveyor belt comprises a Lycra coating.

[0072] In some embodiments, the conveyor device is configured for continuous movement of the solid phase material. In some embodiments, the conveyor device is configured for continuous movement of the solid phase material with periodic slowing or pausing of the conveyor device. In some embodiments, the conveyor device periodically slows or stops to allow for removal of a faulty' cassette or solid phase material. In some embodiments, the conveyor device periodically slows or stops to allow a reaction at any given station to proceed before going to the next station.

[0073] In some embodiments, the choice of material for the conveyor belt depends on the solid phase material used, the concentration and type of reactants and reagents used, as well as other process parameters of the belt reactor. A variety of appropriate materials for the conveyor belt are known and can be used with the present disclosure. Examples include but are not limited to polystyrene, nylon, polyester, cotton, stainless steel, nitrile rubber, PVC (polyvinyl chloride), PE (polyethylene), PET (polyethylene terephthalate), PP (polypropylene). PU (polyurethane). PTFE (polytetrafluoroethylene). PEEK (poly ether ether ketone), PA (polyamide), PVDF (polyvinylidene fluoride), ECTFE (ethylenechlorotrifluoroethylene), SBR (styrene-butadiene rubber), EPDM (ethylene propylene diene monomer rubber), and TPE (thermoplastic elastomer), and RMV (rubber- modified vinyl). In some embodiments, PEEK is a preferred material for the conveyor belt. In some embodiments, one or more processors are programmed to control the operation of the conveyor device.

[0074] In some embodiments, a station comprises one or more liquid collection zones that collect liquid permeating through the conveyor device. Liquid collection zones may comprise collecting tanks that are attached below the conveyor device, which collect any fluid separating from the solid phase material by gravitational force. The fluid collected in these collection zones can be re-used or discarded. In some embodiments, separation of fluid from the solid phase material is accelerated by application of vacuum to the lower part of the conveyor device. In some embodiments, separation of fluid from the solid phase material is accelerated by application of pressure to the upper part of the conveyor device.

[0075] In some embodiments, the fluid collection zones are configured for re-use of the recovered reactant, reagent, or washing fluid. The number of liquid collection zones within a station is flexible and can vary from station to station.

[0076] In some embodiments, the re-use of recovered reactant, reagent, or washing fluid is conducted in cocurrent or countercurrent fashion. When configured for countercurrent reuse, the fluid delivery device closest to the end of the station is supplied with fresh reagent, reactant, or washing fluid. The fluid recovered in this collection zone is fed to an adjacent fluid delivery device located further away from the end of the station. The fluid recovered in this collection zone is fed to a fluid delivery device located even further from the end of the station. As a result, when recovering and re-using reactant or reagent, the content thereof in the recovered fluid decreases with every recovery / reuse cycle. In the case of recovered and reused washing fluid, the content of contaminant increases with every recovery / reuse cycle.At the same time, the amount of oligomer that has been acted upon by the reactant, reagent, or washing fluid increases the further the solid phase material proceeds within the station.

[0077] When configured for cocurrent reuse, the fluid delivery device furthest from the end of the station is supplied with fresh reagent, reactant, or washing fluid. The fluid recovered in this collection zone is fed to an adjacent fluid delivery device located closer to the end of the station. The fluid recovered in this collection zone is fed to a fluid delivery device located even closer to the end of the station.

[0078] In some embodiments, the number of recycling steps within a station corresponds to the number of distinct collection zones within the station and can be adjusted according to the specific requirements of each station.

[0079] In some embodiments, a pressure gradient is used at one or more stations to facilitate the flow of reactant, reagent, or washing fluid through the conveyor device. This may be achieved, e.g., by applying vacuum to the area underneath the conveyor device and / or through applying positive pressure to the area above the convey or device.

[0080] In some embodiments, the time the solid phase material is exposed to the reactant, reagent, or washing fluid can be adjusted. If longer exposure is desired, a pressure gradient may be applied gradually and / or only to a later part of the station, increasing the time the solid phase material is exposed to the reactant, reagent, or washing fluid that is dispensed at that station. In some embodiments, the time the solid phase material is exposed to the reactant, reagent, or washing fluid may also be adjusted by increasing the space between stations. Increasing the space between stations leads to an increased time the solid phase material on the conveyor device takes to reach the next station. Consequently, the solid phase material may be exposed to the reactant, reagent, or washing fluid dispensed at that station for a longer period of time. Increasing the length of stations where slow reactions are carried out may thus facilitate greater reaction completion and / or increase the purity of the synthesized oligomer.

[0081] In some embodiments, a process control device monitors and controls some or all parts of the belt reactor that include but are not limited to devices for creating a pressure gradient, flow meters, valves, conveyor devices, fluid delivery devices, and analytical devices. In some embodiments, the process control device comprises software to monitor and / or control the delivery7of reagents and reactants and / or the movement of the solid phase material. In some embodiments, the software is run on a computing device in electoral communication with the belt reactor apparatus.

[0082] In some embodiments, analytical devices are used to monitor the product or intermediate of a station. In some embodiments, the analytical devices are in electrical communication with the process control device. Examples include but are not limited to UV spectrometer, IR spectrometer, ATR spectrometer, Raman spectrometer, NMR spectrometer, LCMS, mass spectrometer, light monitor, pH meter, nephelometer, turbidity7meter, refractometer, temperature monitor and / or devices configured to measure conductivity or to count particles.

[0083] In some embodiments, analytical devices can act as quality control during oligomer synthesis, e.g., by directly testing material as it passes by the analytical device or by extracting a sample for testing by the analytical device. In the case that a faulty or unsatisfactory product or intermediate is detected, some or all of the faulty material can be discharged into a separate waste tank. In some embodiments, this system may allow for all or a subset of material to be discarded without the need to waste an entire batch of material.

[0084] In some embodiments, the reactant and / or reagent recovered in a collection zone is analyzed by one or more analytical devices to determine progress of the reaction. If reaction progress is below a certain threshold, the conveyor speed is deaccelerated, and / or more reactant or reagent is dispensed to allow for increased exposure of the solid phase material to the reactant and / or reagent. In some embodiments, the pressure gradient at that station is adjusted if reaction progress is determined to be below a certain threshold, thus allowing an increased exposure time.

[0085] In some embodiments, the oligomer produced in a station is analyzed by one or more analytical devices. If the product quality7falls below a certain threshold, an alert is sent to process control and part of, or the entire sample is discarded into a waste tank.

[0086] In some embodiments, the washing fluid recovered in a collection zone is analyzed by one or more analytical devices to determine presence and levels of residual reactants, reagents, and / or impurities. If the levels of residual reactants and / or reagents or impurities is above a certain threshold, the conveyor speed is deaccelerated, and more washing fluid is dispensed to allow for an extended and more rigorous washing step.Sequence of Steps

[0087] In some embodiments, solid phase material that comprises an already attached nucleotide or amino acid is used. In some embodiments, solid phase material without a preattached nucleotide or amino acid is used. In some embodiments, the solid phase material is supplied directly onto the conveyor device, e.g., conveyer belt, for oligomer synthesis. Theconveyor device, e.g., conveyor belt, continuously transports the solid phase material through all modules and stations therein. In some embodiments, the solid phase material is transported through a conveyor device without the need for a conveyor belt.

[0088] In some embodiments, in case a solid phase material with pre-attached nucleotide or amino acid is used, the solid phase material is deposited onto the conveyor device and transported to the first module. In some embodiments, the first module commences with a deprotection station, where one or more reagents are dispensed from a fluid delivery device onto the solid phase material. The solid phase material remains exposed to one or more reagents for a period of time, e.g, until excess fluid is drained through the conveyor device.

[0089] In some embodiments, prior to additional steps, e.g., when a solid phase material is pre-attached to nucleotides or amino acids, the solid phase material leaves the deprotection station and enters a supplemental washing station, where washing fluid is dispensed from a fluid delivery' device onto the solid phase material. The solid phase material remains exposed to the washing fluid for a period of time, e.g., until excess fluid is drained through the conveyor device.

[0090] In some embodiments, where pre-attached nucleotide or amino acid is used, the solid phase material leaves the supplemental washing station and enters a coupling station, where reactant, and optionally, one or more reagents are dispensed from a fluid delivery' device onto the solid phase material. The solid phase material remains exposed to the reactant and the reagent (if used) for a period of time, e.g.. until excess fluid is drained through the conveyor device.

[0091] In some embodiments, solid phase material is supplied directly onto the convey or device for oligomer synthesis. The convey or device continuously transports the solid phase material through all modules and stations therein.

[0092] In some embodiments, where a solid phase material without pre-attached nucleotide or amino acid is used, after deposition onto the conveyor device, the solid phase material may be transported to a coupling station, where reactant and, optionally, one or more reagents are dispensed from a fluid delivery’ device onto the solid phase material. The solid phase material remains exposed to the reactant and the reagent (if used) for some time, e.g. until excess fluid is drained through the conveyor device.

[0093] In some embodiments, the solid phase material leaves the coupling station and enters a first washing station, where washing fluid is dispensed from a fluid delivery' device onto the solid phase material. The solid phase material remains exposed to the washing fluid for a period of time, e.g, until excess fluid is drained through the conveyor device.

[0094] In some embodiments, the solid phase material leaves the first washing station and enters an oxidation station, where oxidant is dispensed from a fluid delivery device onto the solid phase material. The solid phase material remains exposed to the oxidant for a period of time, e.g., until excess fluid is drained through the conveyor device.

[0095] In some embodiments, the solid phase material leaves the oxidation station and enters a second washing station, where washing fluid is dispensed from a fluid delivery device onto the solid phase material. The solid phase material remains exposed to the washing fluid for a period of time, e.g., until excess fluid is drained through the conveyor device.

[0096] In some embodiments, the solid phase material leaves the second washing station and enters a capping station, where a mixture of capping agent and catalyst is dispensed from a fluid delivery’ device onto the solid phase material. In some embodiments, the solid phase material leaves the second washing station and enters a capping station, where a capping agent is dispensed from a fluid delivery device onto the solid phase material. The solid phase material remains exposed to the capping mixture for a period of time, e.g., until excess fluid is drained through the conveyor device.

[0097] In some embodiments, the solid phase material leaves the capping station and enters a third washing station, where washing fluid is dispensed from a fluid delivery device onto the solid phase material. The solid phase material remains exposed to the washing fluid for a period of time, e.g., until excess fluid is drained through the convey or device and collected in a collecting tank. In some embodiments, the end of the third washing station marks the end of the first module, at the end of which the solid phase material is collected in a buffer tank or moves directly to a second module.

[0098] In some embodiments, the solid phase material from the buffer tank is supplied to a reaction vessel containing reagent for the deprotection of the newly added nucleotide or amino acid. The solid phase material remains exposed to the deprotection reagent for some time until it is supplied onto a conveyor device and transported to the next module.

[0099] In some embodiments, one or more additional stations may be positioned before, interspersed between or placed after the steps outlined above, such that additional or alternate washing fluid, reactant, and / or reagent may be delivered to the solid phase material.

[0100] In some embodiments, the belt reactor includes one or more stations for the purpose of cleavage of the produced oligomer from the solid phase material. Cleavage may be achieved in a variety of ways, depending on the linker used. Examples include but are not limited to chemical cleavage reactions, change in pH. change in temperature, or application of radiation.

[0101] In some embodiments, the synthesized oligomer is subjected to further purification, separation, and / or other processes after cleavage from the solid phase material. A variety of methods for post-synthesis processing are known in the art and may be used in this step.

[0102] In some embodiments, parameters controlling reaction times and / or output from a belt reactor for a given solid phase material include belt speed, solid phase height, and / or solid phase width. As such, a scale-up in operation can be achieved in a variety of ways, such as but not limited to increasing the belt speed, increasing the solid phase material loading on the belt, and / or wider belt width. The values for any of these variables may depend on the desired oligomer, their purity, as well as other process parameters.

[0103] Without being bound by theory, the present disclosure provides several advantages over traditional solid phase synthesizers such as higher material throughput, higher conversion and yield, higher purity, better product quality, lower cost of production, better manufacture flexibility', reduced amount of reactant, reagent, or solvent used, faster reaction kinetics, easier scalability', less failure sequences, and lower risk of a mistake ruining a batch of product. In some embodiments, the recycling and reuse of reactant, reagent, and washing fluid provides for a decreased amount of reactant, reagent, and washing fluid required and facilitates higher conversion and yield of the desired oligomer. In some embodiments, a continuous production model for solid phase oligomer synthesis facilitates reduction of production cost and increases manufacturing speed. In some embodiments, the use of liquidspecific feeding lines obviates the need for a cleaning step for every process step, thus reducing production time and loss of liquid. In some embodiments, the adaption of a fluid delivery7system obviates the need for high precision pumps for dosing liquids, thus reducing production cost of the oligomer. In some embodiments, the use of a functionalized solid phase belt as solid phase material reduces the need of the reactant, reagent, or washing fluid to diffuse through the particle’s pores of the solid phase material. Without being bound by theory, this may provide several advantages over traditional solid phase synthesizers such as lower cost of operation, facilitating a higher oligomer loading density, as well as enabling faster reactions, higher throughput, better product quality, and / or less failure sequences. In some embodiments, the use of a solid phase belt provides several advantages over alternative solid phase materials. In some embodiments, the use of a solid phase belt is more cost effective than other solid phase materials. In some embodiments, the use of a solid phase belt eliminates in part or in whole issues arising from a pressure drop in the existing solid phase material. In some embodiments, the use of a solid phase belt eliminates in part or in whole issues arising from pore diffusion. In some embodiments, the use of a solid phase belteliminates issues arising from swelling of the solid phase material in certain solvents. In some embodiments, the use of a solid phase belt provides a higher loading of oligomer compared to other solid phase materials.

[0104] In some embodiments, the belt reactor is configured for the synthesis of peptides using the same principles and steps as for the synthesis of oligonucleotides but using different reactants, reagents and / or washing fluids. For example, the process of solid phase peptide synthesis may comprise the following sequence of steps: (I) Coupling an N-protected amino acid to a suitable solid phase material. (2) Deprotecting the a-amino group of said amino acid using suitable reagents, depending on the protecting group used. (3) Contacting the solidbound amino acid with a mixture of N-protected amino acid reactant and reagents suitable for enabling formation of a peptide bond between the a-amino terminus of the solid-bound amino acid and the carboxyl terminus of the amino acid reactant. (4) Repeating steps 1-3 until the desired peptide length has been reached. (5) Optionally, a step of capping is included after the coupling step to block the ends of unreacted amino acids from reacting in the next coupling step. (6) Washing steps can be incorporated into the sequence to remove residual reactant or reagent from the solid phase material as needed. (7) A step of cleaving the peptide from the solid phase material and, optionally, carrying out steps of purification and / or drying may be included. Each of the steps can be carried out at a station of the belt reactor. Accordingly, solid phase materials, linkers, reactants, reagents, and / or washing fluids can be used, and the stations and process parameters can be adjusted to accommodate peptide synthesis (e.g., to space out stations to accommodate different reaction times for the steps in peptide synthesis).Solid Phase Material

[0105] The solid phase material used herein can comprise different materials, forms, and configurations.

[0106] In some embodiments, the solid phase material is a commercially available solid support such as Nittophase® or Tentagel®. There are, however, other suitable solid phase materials known in the art that can be used with the present disclosure. Examples include but are not limited to a macroporous polystyrene polymer, a crosslinked polystyrene polymer, and a controlled pore glass. In some embodiments, a preferable solid phase material comprises functional hydroxyl groups in high loading. The use of a macroporous polystyrene resin as solid phase material may be preferrable in some instances. In some embodiments, the solid phase material comprises polymeric materials in form of particles with inner pores. In some embodiments, the solid phase material comprises glass in form of particles with innerpores. In some embodiments, the solid phase material comprises carbon in form of particles with inner pores.

[0107] In some embodiments, the solid phase material is made from one or more of polymers, resins, controlled porous glass, graphene, graphene oxide, and glass. In some embodiments, the solid phase material is of polymeric material. In some embodiments, the solid phase material is made of a resin. In some embodiments, the solid phase material is made of graphene. In some embodiments, the solid phase material is made of graphene oxide. In some embodiments, the solid phase material is made of reduced graphene oxide. In some embodiments, the solid phase material is made of glass.

[0108] In some embodiments, the solid phase material comprises a solid phase belt. In some embodiments, the solid phase material comprises a nonwoven fiber. In some embodiments, the preferred material the nonwoven fiber is glass. In some embodiments, the preferred material the nonwoven fiber is polystyrene. In some embodiments, the preferred fiber diameter of the nonw oven fiber is 1 to 10 pm. In some embodiments, the preferred fiber diameter of the nonwoven fiber is 0.2 to 1 pm. Such fibers may be produced through electrospinning (with or without air blowing) or centrifugal spinning (Bhardwaj et al., “Electrospinning: A fascinating fiber fabrication technique” Biotechnology Advances 2010, 28(3), 325-347; Agarwal et al., “Use of electrospinning technique for biomedical applications” Polymer 2008, 49(26). 5603-5621).

[0109] In some embodiments, the solid phase material is treated to increase or add binding capabilities to one or more of an amino acid, peptide, nucleotide, and oligonucleotide. In some embodiments, the solid phase material is modified to allow for solid phase oligomer synthesis. In some embodiments, the treating of solid phase material increases the presence of hydroxy groups on the solid phase material. In some embodiments, the treating of solid phase material is achieved through one or more of plasma treatment, radiation, coating, UV radiation, chemical grafting, and annealing. In some embodiments, the treating of solid phase material is achieved through plasma treatment. In some embodiments, the treating of solid phase material is achieved through radiation. In some embodiments, the treating of solid phase material is achieved through coating. In some embodiments, the treating of solid phase material is achieved through chemical grafting. In some embodiments, the treating of solid phase material is achieved through annealing. In some embodiments, the treating of solid phase material through radiation comprises ultraviolet radiation.

[0110] In some embodiments, the oligomer is directly attached to the solid phase material. In some embodiments, the oligomer is attached to the solid phase material via a linker. A varietyof linkers are known in the art that can be used in a belt reactor. Examples include but are not limited to a Wang linker, Rink linker, Sheppard linker, phenylfluorenyl linker, oxazolidine linker, acetal linker, methacrylic linker, DDE linker, vinyl sulphone linker, REM linker, sulphone linker, photolabile linker, arylsulphonate linker, and cysteine linker.[OHl] In some embodiments, the solid phase material is in a form selected from particles, fibers, woven fibers, nonwovens, felts, textiles, gels, flat surfaces, tapes, membranes, foams, extrudates, and extruded materials. In some embodiments, the solid phase material comprises fibers. In some embodiments, the solid phase material comprises fibers with a preferred width of 0.2 to 10 pm. In some embodiments, the solid phase material comprises woven fibers. In some embodiments, the solid phase material comprises nonwovens. In some embodiments, the solid phase material comprises felts. In some embodiments, the solid phase material comprises textiles. In some embodiments, the solid phase material comprises gels. In some embodiments, the solid phase material comprises flat surfaces. In some embodiments, the solid phase material comprises tapes. In some embodiments, the solid phase material comprises membranes. In some embodiments, the solid phase material comprises foams. In some embodiments, the solid phase material comprises extruded materials. In some embodiments, the solid phase material comprises a mixture of two or more of particles, fibers, woven fibers, nonwovens, felts, textiles, gels, flat surfaces, tapes, membranes, foams, and extruded materials.

[0112] In some embodiments, the solid phase material is a nonwoven material. In some embodiments, the nonwoven material is modified with linkers known in the art to allow for solid phase oligomer synthesis. In some embodiments, the linkers are located in pores of the solid phase material. In some embodiments, the linkers are located on the surface of the fiber, resulting in an increased mass transfer of the reagent, reactant, or washing fluid through the solid phase material due to elimination of pore diffusion. In some embodiments, use of a nonwoven material may reduce reaction times, and / or a higher throughput may be achieved. In some embodiments, the cost of nonwoven materials is lower than that of other solid phase materials.

[0113] Several nonwoven materials can be used with the present disclosure. Examples include but are not limited to a polystyrene, nylon, polyester, cotton, nitrile rubber, PVC (polyvinyl chloride), PE (polyethylene), PET (polyethylene terephthalate), PP (polypropylene), PU (polyurethane). PTFE (polytetrafluoroethylene), PEEK (polyether ether ketone), PA (polyamide), PVDF (polyvinylidene fluoride), ECTFE (ethylenechlorotrifluoroethylene), SBR (styrene-butadiene rubber), EPDM (ethylenepropylene diene monomer rubber), TPE (thermoplastic elastomer), and RMV (rubber- modified vinyl).

[0114] In some embodiments, the solid phase fibers, woven fibers, nonwovens, felts, textiles, gels, flat surfaces, tapes, membranes, foams, or extruded materials are comprised within the flexible section of a cassette, which is transported on the conveyor device. In some embodiments the flexible section of the cassette is configured in a single layer by the rigid section of the cassette. In some embodiments the flexible section of the cassette is configured in two or more layers by the rigid section of the cassette. In some embodiments, the two or more layers of the flexible section of the cassette are of the same solid phase material type. In some embodiments, the two or more layers of the flexible section of the cassette are different solid phase materials.

[0115] In some embodiments, the multiple layers of flexible section of the cassette are flat, folded, rolled, spiral, star-shaped, stacked, or a mixture of any of the foregoing. In some embodiments, the arrangement of the flexible section of the cassette increases the loading of oligomers per surface area of conveyor device. In some embodiments, the arrangement of the flexible section of the cassette increases the density of oligomer chains per cassette.

[0116] In some embodiments, the use of cassettes in solid phase material allows for a quasi- continuous manufacturing. In some embodiments, the use of cassettes allows for a continuous manufacturing of oligomers while retaining the benefit of discrete belt units. In some embodiments, the use of cassettes in solid-phase manufacturing allows specific cassettes to be discarded during operation. In some embodiments, the use of cassettes in solid-phase manufacturing allows for select cassettes that comprise faulty' solid phase material to be discarded without affecting the remaining solid phase materials of other cassettes.

[0117] In some embodiments, the cassette is configured to seal the flexible section from the outside. In some embodiments, sealing of the flexible section of the cassette allows temporal or permanent storage of the solid-bound oligomers. In some embodiments, sealing of the cassette prevents exposure of the solid-bound oligomers to moisture, oxygen, light, or other undesirable conditions that may negatively impact the solid-bound oligomers.Manufacturing Reagents and Reactants

[0118] In some embodiments, one or more stations are configured to chemically modify the last monomer that was added to an oligomer sequence. In the case of oligonucleotide synthesis, examples include but are not limited to removing a protecting group from, linking a new monomer to, oxidizing a phosphite to a phosphate moiety in, converting a phosphitetriester to a phosphorothioate in, and capping residual functional groups of the nucleotide that was last added to the oligonucleotide sequence. In the case of peptide synthesis, examples include but are not limited to removing a protecting group from, linking a new monomer to, and capping residual functional groups of the amino acid that was last added to the peptide sequence.

[0119] In some embodiments, a deprotection step comprises supplying solid phase material to a deprotection station, where a protecting group is removed from a solid-bound monomer or oligomer. A variety of protecting groups and different deprotection conditions are available in the art. A protecting group and corresponding deprotection conditions are chosen based on the nucleotides or amino acids involved and other process parameters. Examples of protecting groups for use in oligonucleotide synthesis include but are not limited to a trityl and DMT protecting group. Examples of protecting groups for use in peptide synthesis include but are not limited to Bn, Boc, BOM, TBS, TBDPS, Cbz, Fmoc, DMB, SEM, Aloe, tert-butyl, DMT, trityl, and benzoyl protecting group. The use of DMT as protecting group for oligonucleotide synthesis and Fmoc as protecting group for peptide synthesis may be preferrable in some instances.

[0120] In some embodiments, a deprotection step comprises dispensing one or more reagents onto the solid phase material to cleave a protecting group from the solid-bound oligomer. A reagent used for deprotection may be dispensed in neat form or as a solution in an appropriate solvent. The choice of deprotection reagent, amount, and concentration may depend on the specific protecting group, the nucleotides or amino acids to be treated, as well as other process parameters and may be adjusted as needed. Several reactants are known in the art and may be used in this step. Examples of reagents for cleavage of acid-labile protecting groups include but are not limited to acetic acid, dichloroacetic acid, trichloroacetic acid, trifluoroacetic acid, CSA, PTSA, mineral acids, and other applicable organic acids. The use of trifluoroacetic acid may be preferrable in some instances. Examples of reagents for cleavage of base-labile protecting groups include but are not limited to pyridine, triethylamine, methylamine, imidazole, ammonia, and other applicable organic bases. The use of imidazole may be preferrable in some instances.

[0121] In some embodiments, a coupling step comprises dispensing reactant and / or one or more reagents onto the solid phase material to grow the nucleotide chain. A reagent may be dispensed in neat form or as a solution in an appropriate solvent. Several reactants are known in the art and may be used in this step. In the case of oligonucleotide synthesis, examplesinclude but are not limited to phosphoramidite, phosphotriester, and H-phosphonate. The use of phosphoramidite reactants may be preferrable in some instances.

[0122] In some embodiments, a coupling step comprises dispensing reactant and, optionally, one or more reagents onto the solid phase material to grow the peptide chain. A reactant and / or reagent may be dispensed in neat form or as a solution in an appropriate solvent. Several reactants are known in the art and may be used in this step. Examples include but are not limited to N-protected amino acids and C-protected amino acids. Depending on which terminus is anchored to the solid phase, the C-terminus, the N-terminus, and / or optionally one or more side chain functional groups may be protected with appropriate protecting groups to facilitate selective reaction at the unprotected site. Examples of reagents for coupling amino acids include but are not limited to COMU, DCC. DIC, EDC. HBTU, TBTU, HATU, TATU, BOP, PyBOP, HOAt, and HOBt. The use of T3P in combination with COMU may be preferrable in some instances.

[0123] In some embodiments, one or more of the reagents employed in the coupling step comprise a catalyst. A catalyst may be dispensed in neat form or as a solution in an appropriate solvent. In the case of solid phase oligonucleotide synthesis, examples of catalysts include but are not limited to IH-tetrazole, 5-(4-nitrophenyl)-lH-tetrazole, 5-(bis- 3,5-trifluoromethylphenyl)-lH-tetrazole, 5-methylthio-lH-tetrazole, 5-ethylthio-lH-tetrazole, 5-benzylthio-lH-tetrazole, 5-mercapto-tetrazole, 1 -hydroxy -benzotriazole, 3-nitro-lH-l,2,4- triazole, pyridinium trifluoroacetate, imidazolium triflate. imidazolium perchlorate, imidazolium tetrafluoroborate, N-(methyl)imidazolium triflate, N-(phenyl)imidazolium triflate, N-(phenyl)imidazolium perchlorate, N-(phenyl)imidazolium tetrafluoroborate, N-(p- acetylphenyl)imidazolium triflate, 2-(phenyl)imidazolium triflate, 4-(methyl)imidazolium triflate. 4-(methyl)imidazolium tosylate. 4-(methyl)imidazolium trifluoroacetate, 4- (phenyl)imidazohum triflate, 4-(phenyl)imidazolium trifluoroacetate, benzimidazolium triflate, benzimidazolium tetrafluoroborate, N-(methyl)benzimidazolium triflate, 2- (phenyl)benzimidazolium triflate, 2-(phenyl)benzimidazolium perchlorate, diisopropylammonium tetrazolide, and 4,5-dicyanoimidazole. The use of 5-ethylthio-lH- tetrazole may be preferrable in some instances.

[0124] In some embodiments, a washing step comprises removing residual reagent or reactant from the solid phase material by washing with an appropriate liquid. The liquid can comprise a single solvent or a mixture of solvents. Several solvents are known in the art and may be used in this step. Examples include but are not limited to acetonitrile, dichloromethane, dichloroethane, chloroform, dimethoxyethane, N,N-dimethylformamide,N,N’ -dimethylacetamide, N-methyl-2 -pyrrolidone, N-formylmorpholine, N- butylpyrrolidinone, cyclo pentyl methyl ether, methyl tert-butyl ether, diethyl ether, tetrahydrofuran, toluene, 2-methyltetrahydrofuran, ethyl acetate, dimethyl carbonate, dioxane, methanol, ethanol, iso-propanol, water, and dimethyl sulfoxide. The use of acetonitrile may be preferrable in some instances.

[0125] In some embodiments, an oxidation step comprises dispensing one or more reagents onto the solid phase material to oxidize a phosphite to a phosphate moiety. A reagent for oxidation may be dispensed in neat form or as a solution in an appropriate solvent. Several oxidants are known in the art and may be used in this step. Examples include but are not limited to iodine, tert-butyl hydroperoxide, and 10-camphorsulfonyl-oxaziridine. The use of iodine may be preferrable in some instances.

[0126] In some embodiments, a thiolation step comprises dispensing one or more reagents onto the solid phase material to achieve thiolation. A reagent for thiolation may be dispensed in neat form or as a solution in an appropriate solvent. Several thiolation agents are known in the art and may be used in this step. Examples include but are not limited to DDTT, 3H-1,2- benzodithiol-3-one 1,1-dioxide, xanthane hydnde and N,N.N'N'-tetraethylthiuram disulfide. The use of xanthane hydride may be preferrable in some instances.

[0127] In some embodiments, a capping step comprises dispensing one or more reagents onto the solid phase material to achieve capping of unreacted functional groups, hindering them from further reaction. A capping reagent may be dispensed in neat form or as a solution in an appropriate solvent. Several capping agents are known in the art and may be used in this step. Examples include but are not limited to acetic anhydride, acetyl chloride, propionic anhydride, isobutyric anhydride, phenoxy acetic anhydride, and / c / 7-biitylphenoxyacetic anhydride. The use of acetic anhydride may be preferrable in some instances.

[0128] In some embodiments, one or more of the reagents in the capping step comprise a catalyst. A catalyst may be provided in conjunction with another capping reagent, either or both of which may be dispensed in neat form or as a solution in an appropriate solvent. Several catalysts and capping reagents are known in the art and may be used in this step. Examples of catalysts include but are not limited to N-methylimidazole, 4- dimethylaminopyridine, and pyridine. Examples of capping reagents include but are not limited to acetic anhydride, acetyd chloride, propionic anhydride, isobutyric anhydride, phenoxy acetic anhydride, and te / 7-butylphenoxyacetic anhydride. The use of NMI and acetic anhydride may be preferrable in some instances.Exemplary Arrangement

[0129] Fig. 1 shows an overview of an exemplary setup of a belt reactor. The depicted belt reactor comprises three modules (1), and each module comprises seven stations. In a reaction vessel (2) containing solid phase particles (3), a deprotection step is carried out, and, upon completion of the reaction, the solid phase material is deposited to the first module. Excess reactant, reagent, or washing fluid is collected in a waste tank (4), and buffer tanks (5) are used to collect the solid phase particles in between modules. Feeding tanks (6-16) supplyreagent, reactant, or washing fluid to the respective stations through pipes, tubes, or functional equivalents thereof (14). For the sake of clarity, only select pipes, tubes, or functional equivalents thereof are depicted. Feeding tank (8) supplies a solution comprising a reactant that effects incorporation of C into the oligomer; feeding tank (9) supplies a solution comprising a reactant that effects incorporation of G into the oligomer; and feeding tank (10) supplies a solution comprising a reactant that effects incorporation of T into the oligomer. Process control (17) monitors and controls operation of all substantive parts of the belt reactor, such as opening / closing of valves (18), conveyor device speed, and dispensing of fluid.

[0130] Fig. 2 shows an example of a module comprising seven stations wherein each station is responsible for carry ing out a specific step in the oligomer synthesis (with the respective step indicated in the box above the station). In a reaction vessel (1) a deprotection step is carried out. Upon completion, the solid phase particles (2) are deposited onto a conveyor device (3), which moves the solid phase particles through each station. At each station, a fluid delivery7device (4-10) dispenses reagent, reactant, or washing fluid onto the solid phase material. Vacuum is applied to the underside of the conveyor device to increase the flow of fluid through the solid phase particles. One or more collection zones (11) are present for the recovery of reactant, reagent, or washing fluid.

[0131] Fig. 3 shows an example of a station. The solid phase particles (1) are moved through the station via a conveyor device (2). A feeding tank (3) supplies a reactant, a reagent, and / or washing fluid to a fluid delivery device (4), which dispenses it onto the solid phase particles. Excess reactant, reagent, or washing fluid is recovered by liquid collection zones (5-7) and fed back to a fluid delivery device.

[0132] Fig. 4 shows an example of a module in a rotary- configuration comprising seven stations wherein each station is responsible for carry ing out a specific step in the oligomer synthesis. Solid phase material (1) is deposited onto a conveyor device (2) on a moving rotary cylinder (3) from a buffer tank (4) and moved through each station by rotation (5) of therotary cylinder. At each station, a fluid delivery device (6-12) dispenses reagent, reactant, or washing fluid onto the solid phase material. Vacuum is applied to the underside of the conveyor device to increase the flow of fluid through the solid phase material. One or more collection zones (13) are present for the recovery of reactant, reagent, or washing fluid. At the end of the module, the solid phase material that has been acted upon (14) is removed from the conveyor device.

[0133] Fig. 5 shows an overview of an exemplary setup of a belt reactor operating with a solid phase belt as solid phase material. The depicted belt reactor comprises three modules (1), and each module comprises seven stations. The solid phase belt (2) is unrolled from a reel (3) and fed to the first station of the first module. Then, the individual stations act on the solid phase belt to grow the oligomer chain. Excess reactant, reagent, or washing fluid is collected in a waste tank (4). Feeding tanks (5-14) supply reagent, reactant, or washing fluid to the respective stations through pipes, tubes, or functional equivalents thereof (15). For the sake of clarity7, only a subset of the pipes, tubes, or functional equivalents thereof that may be used in the disclosed methods are depicted in Fig. 5. Feeding tank (7) supplies a solution comprising a reactant that effects incorporation of C into the oligomer; feeding tank (8) supplies a solution comprising a reactant that effects incorporation of G into the oligomer; and feeding tank (9) supplies a solution comprising a reactant that effects incorporation of T into the oligomer. Process control (16) monitors and controls operation of all substantive parts of the belt reactor, such as opening / closing of valves (17). conveyor device speed, and dispensing of fluid.

[0134] Fig. 6 shows an example of a single module of a belt reactor operating with a solid phase belt (1) as solid phase material. The solid phase belt comprises chemical modifications on its surface. A linker (2) is configured to connect the growing oligomer (3) to the solid phase belt. The solid phase belt is unrolled from its reel (4) and deposited onto the conveyor device (5). The parts of the belt reactor employing a solid phase belt as solid phase material may be substantially similar to that outlined in Fig. 1-3, except for differences stemming from the different solid phase material.

[0135] Fig. 7 shows an example of a single station of a belt reactor operating with a solid phase belt as solid phase material. The solid phase belt (1) is moved through the station via a conveyor device (2). A feeding tank (3) supplies a reactant, a reagent, and / or washing fluid to a fluid delivery device (4), which dispenses it onto the solid phase belt. Excess reactant, reagent, or washing fluid is recovered by liquid collection zones (5-7) and fed back to a fluid delivery device.

[0136] Fig. 8 shows an example of a single module with eight horizontally arranged stations within a belt reactor, operating with cassettes that comprise a flexible section configured for coupling to oligomers (1) and a rigid frame (2). The working principle of all substantive parts of the belt reactor is substantially similar to that outlined in Fig. 1-3, except for differences stemming from the use of cassettes.

[0137] Fig. 9 shows six examples of cassettes: (a) a single sheet of a flexible band (1) arranged with a rigid section (2) to form a single cassette; (b) several sheets of flexible band of the same material (3) arranged with a rigid section (2) to form a single cassette; (c) several sheets of flexible band arranged with a rigid section (2) to form a single cassette, wherein the material of flexible band (4) differs from that of the other flexible band (5); (d) a single sheet of a flexible band (6) arranged with a rigid section (2) to form a single cassette; (e) several sheets of flexible band (7) arranged in a folded manner with a rigid section (2) to form a single cassette; and (f) several sheets of flexible band arranged in a folded manner with a rigid section (2) to form a single cassette, wherein the material of one flexible band (8) differs from that of the other flexible band (9).Examples

[0138] The following examples are presented to illustrate possible embodiments and are not meant to be limiting of the scope.Example 1 - Oligonucleotide Synthesis

[0139] An example of a belt reactor as disclosed herein is described below for the synthesis of an oligonucleotide. An oligonucleotide is produced using the apparatus and steps as described herein. A process control device is configured to control the movement of the solid phase material on the conveyor device through the stations and delivery of the appropriate amounts of reactants, reagents, and washing fluid to the solid phase material in a sequence to synthesize the desired oligonucleotide. Reactants, reagents, and w ashing fluids are recovered and reused in a countercurrent fashion at all stations.

[0140] (1) Solid phase material comprising a macroporous polystyrene resin is deposited onto a permeable conveyor belt and transported to a first module for linking a nucleotide to the solid phase material. As the solid phase material continuously moves through the first station, a fluid comprising a solution of nucleoside phos phorami dite and 5-(4-nitrophenyl)- IH-tetrazole in acetonitrile is dispensed from a spray nozzle onto the solid phase material on the conveyor belt. A vacuum is applied to the underside of the conveyor belt to increase the flow7of the fluid through the solid phase material. The liquid recovered this w ay is fed toanother spray nozzle within the same station, dispensing the reused liquid onto an earlier portion of the solid phase material.

[0141] (2) As the solid phase material leaves the coupling station along the conveyor belt and enters the first washing station, a washing fluid comprising acetonitrile is dispensed onto the solid phase material, removing residual reactant and reagent.

[0142] (3) Entering the oxidation station, a fluid comprising a solution of iodine, pyridine, and water in acetonitrile is dispensed from a spray nozzle onto the solid phase material, oxidizing the phosphite to a phosphate.

[0143] (4) After leaving the oxidation station, the solid phase on the conveyor belt enters a washing station. A washing fluid comprising acetonitrile is dispensed onto the solid phase material, removing residual reagent.

[0144] (5) The solid phase material enters the capping station, where a fluid comprising a solution of acetic anhydride and N-methylimidazole in acetonitrile is dispensed onto the solid phase material.

[0145] (6) The solid phase material then moves along the conveyor belt to a washing station. A washing fluid comprising acetonitrile is dispensed onto the solid phase material, removing residual reagent.

[0146] (7) The solid phase material leaves the first module and is collected in a buffer tank.

[0147] (8) A solution of dichloroacetic acid in acetonitrile is added to the tank, removing the DMT protecting group from the 5 ’-oxygen on the nucleotide added in step 1.

[0148] (9) The solid phase material is supplied to a new module that starts with a washing station to remove residual reagent from the previous step. The solid-bound oligonucleotide is further grown by subjecting it to steps 1-9 of either the same or different modules. Once the desired oligonucleotide sequence has been prepared, the oligonucleotide is cleaved from the solid support and may be subjected to further post-synthesis procedures.Example 2 - Peptide Synthesis

[0149] An example of a belt reactor as disclosed herein is described below for the synthesis of a peptide. A peptide is produced using the apparatus and steps as described herein. A process control device is configured to control the movement of the solid phase material on the conveyor device through the stations and delivery of the appropriate amounts of reactants, reagents, and washing fluid to the solid phase material in a sequence to synthesize the desired peptide. Use of the countercurrent principle described previously is applied to all stations.

[0150] (1) Solid phase material comprising a Rink amide resin with a pre-attached amino acid is deposited onto a permeable conveyor belt and transported to first module for linking aN-protected amino acid to the solid phase material. As the solid phase material continuously moves through the first station, a fluid comprising a solution of N-Fmoc-protected amino acid, EDC, and HOBt in DMF is dispensed from a spray nozzle onto the solid phase material on the conveyor belt. A vacuum is applied to the underside of the conveyor belt to increase the flow of the fluid. The liquid recovered this way is fed to another spray nozzle within the same station, dispensing the reused liquid onto an earlier portion of the solid phase material.

[0151] (2) As the solid phase material leaves the coupling station and enters the first washing station, a washing fluid comprising DMF is dispensed onto the solid phase material, removing residual reactant and reagent.

[0152] (3) The solid phase material enters the capping station, where a fluid comprising a solution of acetic anhydride and pyridine in DMF is dispensed onto the solid phase material.

[0153] (4) The solid phase material on the conveyor belt next enters a washing station. A washing fluid comprising DMF is dispensed onto the solid phase material, removing residual reactant and / or reagent.

[0154] (5) The solid phase material leaves the first module and is collected in a buffer tank.

[0155] (6) A solution of piperidine in DMF is added to the tank, removing the Fmoc protecting group from the amine group of the newly attached amino acid.

[0156] (7) The solid phase material is supplied to a new module that starts with a washing station to remove residual piperidine from the previous step. The solid-bound peptide is further grown by subjecting it to steps 1-7 of other modules. Once the desired peptide sequence is realized, the peptide is cleaved off the solid support and subjected to further postsynthesis procedures.Example 3 - Oligonucleotide Synthesis with a Solid Phase Belt

[0157] An oligonucleotide is produced using the apparatus and steps as described herein. A process control device is configured to control the movement of the solid phase material on the conveyor device through the stations and the delivery of the appropriate amounts of reactants, reagents, and washing fluid to the solid phase material in a sequence to synthesize the desired oligonucleotide. Reactants, reagents, and washing fluids are recovered and reused in a countercurrent fashion at all stations. The solid phase material is a solid phase belt, which comprises a polymeric material that is functionalized to enable coupling of oligonucleotides to the solid phase belts. In this example, a solid phase belt without pre-attached nucleotide is used. A permeable filtration belt comprising PEEK acts as a conveyor device, provides additional mechanical support to the solid phase belt, and assists in the transportation thereof through the belt reactor.

[0158] (1) The solid phase belt is unrolled from its stored form and supplied to the first module of the belt reactor. At this first module, a first nucleotide is to be linked to the solid phase belt. As the solid phase belt continuously moves through the first station of the first module, a fluid comprising a solution of nucleoside phosphorami dite and 5-(4-nitrophenyl)- IH-tetrazole in acetonitrile is dispensed from a spray nozzle onto the solid phase belt. A vacuum is applied to the underside of the filtration belt to increase the flow of the fluid through the solid phase belt and filtration belt. The liquid recovered this way is fed to a different spray nozzle within the same station, which dispenses the reused liquid onto the solid phase belt at an earlier section of the first station. As the solid phase belt reaches the end of the first station, the coupling reaction is complete, and the first nucleotide is attached to the solid phase belt.

[0159] (2) As the solid phase belt leaves the coupling station and enters the first washing station, a washing fluid comprising acetonitrile is dispensed onto the solid phase belt, removing residual reactant and reagent. As the solid phase belt reaches the end of the first station, the reactant, reagent, and other impurities from the coupling station have been washed off the solid phase belt. The solid phase belt leaves the first washing station.

[0160] (3) As the solid phase belt enters the oxidation station, a fluid comprising a solution of iodine, pyridine, and water in acetonitrile is dispensed from a spray nozzle onto the solid phase belt, oxidizing the newly created phosphite bond to a phosphate. Upon reaching the end of the oxidation station, the oxidation reaction is gone to completion, and the solid phase belt exits the oxidation station.

[0161] (4) After leaving the oxidation station, the solid phase belt enters a second washing station. Here, a washing fluid comprising acetonitrile is dispensed onto the solid phase material, effectively removing any residual reagent and other impurities from the oxidation station. Once the solid phase belt reaches the end of the second washing station, all impurities have been washed off. The solid phase belt leaves the second washing station.

[0162] (5) The solid phase belt enters the capping station, where a fluid comprising a solution of acetic anhydride and N-methylimidazole in acetonitrile is dispensed onto the solid phase material. As the solid phase belt approaches the end of the capping station, any residual hydroxy groups have been converted to ethyl esters, thus preventing further undesired reaction with reactants and reagents added in future stations. The solid phase belt leaves the capping station.

[0163] (6) The solid phase belt is transported to the third washing station. Upon entry of the third washing station, a washing fluid comprising acetonitrile is dispensed onto the solidphase material, removing residual reagent. Upon reaching the end of the third washing station, all impurities of the previous station have been washed off. The solid phase belt leaves the third washing station.

[0164] (7) The solid phase belt enters the deprotection station, where a solution comprising dichloroacetic acid in acetonitrile is dispensed onto the solid phase belt, removing the DMT protecting group from the 5 '-oxygen of the nucleotide added in step 1. Once the solid phase belt reaches the end of the deprotection station, all protecting groups have been removed from the newly added nucleotide of step 1. The solid phase belt leaves the deprotection station.

[0165] (8) The solid phase belt is transported to the fourth washing station. Upon entry of the fourth washing station, a washing fluid comprising acetonitrile is dispensed onto the solid phase material, removing residual reagent and other impurities. Upon reaching the end of the fourth washing station, all impurities of the previous station have been washed off. The solid phase belt leaves the fourth washing station.

[0166] (9) The solid phase belt leaves the first module and is transported by the filter belt to the next module of the belt reactor.

[0167] (10) The solid-bound oligonucleotide is further grown by subjecting the solid phase belt to steps 1-9 of additional modules of either the same or different ty pe.

[0168] (11) Once the desired oligonucleotide sequence has been prepared, the solid phase belt leaves the last washing station of the last module of the belt reactor and, instead of repeating steps 1 -9, the thus-prepared oligonucleotide is cleaved from the solid support and optionally subjected to further post-synthesis procedures.

Claims

WHAT IS CLAIMED IS:

1. An apparatus for oligomer synthesis, said apparatus comprising: at least one deprotection station; at least one coupling station; optionally, one or more oxidation and / or thiol ati on stations; optionally, one or more capping stations; and optionally, one or more washing stations; wherein the apparatus is configured to act on one or more moveable solid phase materials at each station to produce an oligomer; and wherein each station is at a fixed position in the apparatus and configured to deliver a reagent, a reactant, or a washing fluid to the moveable solid phase material via a fluid delivery device.

2. The apparatus according to claim 1, wherein the at least one deprotection station carries out a step of deprotecting.

3. The apparatus according to claim 1, wherein the at least one coupling station carries out a step of coupling.

4. The apparatus according to claim 1, wherein the one or more oxidation and / or thiolation stations carries out a step of oxidation or thiolation.

5. The apparatus according to claim 1, wherein the one or more capping stations carries out a step of capping.

6. The apparatus according to claim 1, wherein the one or more washing stations carries out a step of washing.

7. The apparatus according to any one of claims 1 to 6, wherein the oligomer is attached to the solid phase material.

8. The apparatus according to any one of claims 1 to 7, wherein the oligomer is attached to the solid phase material via a linker, preferably by solid phase coupling.

9. The apparatus according to any one of claims 1 to 8, wherein the apparatus is configured to act on one or more solid phase materials in parallel.

10. The apparatus according to any one of claims 1 to 9, wherein the one or more fluid delivery devices comprises a spray nozzle, a perforated nozzle, a perforated plate, or an open Pipe.

11. The apparatus according to any one of claim 1 to 10, wherein the one or more solid phase materials are configured to move through the stations via a conveyor device.

12. The apparatus according to any one of claim 1 to 11, wherein the one or more solid phase materials is configured to move stepwise through the stations via a conveyor device.

13. The apparatus according to any one of claims 1 to 12, wherein the conveyor device comprises a conveyor belt.

14. The apparatus according to claim 13, wherein the conveyor belt comprises a permeable material.

15. The apparatus according to any one of claim 1 to 14, wherein the apparatus comprises a pressure gradient across the conveyor belt to increase the flow of the reagent, reactant, or washing fluid through the permeable material.

16. The apparatus according to claim 15, wherein the pressure gradient is achieved by applying vacuum to the underside of the conveyor belt.

17. The apparatus according to claim 15, wherein the pressure gradient is achieved by applying positive pressure to the top face of the conveyor belt.

18. The apparatus according to any one of claims 1 to 17. wherein the apparatus is configured to recover an excess reagent, reactant, or washing fluid.

19. The apparatus according to any one of claim 1 to 18, wherein the excess reagent, reactant, or washing fluid is recovered in a collecting tank after it has passed through the solid phase material.

20. The apparatus according to claim 18 or claim 19, wherein the recovered reagent, the recovered reactant, or the recovered washing fluid is capable of being reused within the same station.

21. The apparatus according to claim 18 or claim 19, wherein the recovered reagent, the recovered reactant, or the recovered washing fluid is capable of being reused in a different station.

22. The apparatus according to any one of claims 18 to 21, wherein the recovered reagent, the recovered reactant, or the recovered washing fluid is configured to feed back to the fluid delivery device.

23. The apparatus according to claim 22, wherein said fluid delivery device is configured to provide for countercurrent reuse of reagents, reactants, and / or washing fluid.

24. The apparatus according to claim 22, wherein said fluid delivery device is configured to provide for cocurrent reuse of reagents, reactants, and / or washing fluid.

25. The apparatus according to any one of claims 1 to 24, wherein one or more stations in a module are disposed at distances from each other selected to provide for different reaction kinetics of the deprotection, coupling, oxidation, thiolation, and capping step.

26. The apparatus according to any one of claim 1 to 25, further comprising a product analytical device.

27. The apparatus according to claim 26, wherein the product analytical device is configured to provide a signal indicating the solid phase material should be retained, partially retained, or discarded.

28. The apparatus according to any one of claim 1 to 27, wherein two or more stations that carry out the same step and employ the same composition of reactant, reagent, and washing fluid are operably linked to a tank containing the reactant, the reagent, or the washing fluid.

29. The apparatus according to any one of claim 1 to 28, wherein the one or more solid phase material is deposited onto the conveyor device.

30. The apparatus according to any one of claim 1 to 29, wherein the one or more solid phase material comprises particles made from one or more of a controlled pore glass material, a crosslinked polystyrene, and a polystyrene.

31. The apparatus according to any one of claim 1 to 30, wherein the one or more solid phase material comprises a solid phase belt.

32. The apparatus according to any one of claims 1 to 31, wherein the solid phase belt comprises one or more materials selected from polymers, carbon, glass, graphene, and graphene oxide.

33. The apparatus according to any one of claim 1 to 32, wherein the one or more solid phase materials has been modified to increase or add presence of functional groups on the solid phase material.

34. The apparatus according to any one of claim 1 to 33, wherein the one or more solid phase materials comprises particles, fibers, woven fibers, nonwoven fibers, nonwovens, felts, textiles, gels, flat surfaces, tapes, membranes, foams, and extrudates.

35. The apparatus according to any one of claim 1 to 34, wherein the one or more solid phase materials comprises a nonwoven material.

36. The apparatus according to claim 35, wherein the nonwoven material comprises polystyrene, nylon, polyester, cotton, nitrile rubber, polyvinyl chloride, polyethylene, polyethylene terephthalate, polypropylene, polyurethane, polytetrafluoroethylene, polyether ether ketone, polyamide, polyvinylidene fluoride, ethylenechlorotrifluoroethylene, styrene-butadiene rubber, ethylene propylene diene monomer rubber, thermoplastic elastomer, and / or rubber-modified vinvl.

37. The apparatus according to any one of claim 1 to 36, wherein the shape of the solid phase material is flat, folded, star-shaped, rolled, or spiral shaped.

38. The apparatus according to claim 34, wherein the fiber, woven fiber, nonwoven, felt, textile, gel, flat surface, tape, membrane, foam, or extrudate is comprised within the flexible section of a cassette.

39. The apparatus according to claim 38, wherein the fiber, woven fiber, nonwoven, felt, textile, gel. flat surface, tape, membrane, foam, or extrudate is arranged in a configuration to increase the surface area of the fiber, woven fiber, nonwoven, felt, textile, gel, flat surface, tape, membrane, foam, or extrudate.

40. The apparatus according to claim 38 or claim 39, wherein the configuration of the fiber, woven fiber, nonwoven, felt, textile, gel, flat surface, tape, membrane, foam, or extrudate is selected from folded, layered, rolled, and spiraled.

41. The apparatus according to claim 8, wherein the linker is configured to release the oligomer from the solid support by one or more chemical cleavage reactions or by a change in pH and / or temperature.

42. The apparatus according to claim 8 or claim 41, wherein the linker comprises a Wang linker, Sheppard linker, Rink linker, phenylfluorenyl linker, oxazolidine linker, acetal linker, methacr lic linker, DDE linker, vinyl sulphone linker, REM linker, sulphone linker, photolabile linker, aryl sulphonate linker, cysteine linker, and / or a traceless linker.

43. The apparatus according to any one of claim 1 to 42, wherein a buffer tank is configured to collect the one or more solid phase materials at the end of a module and feed it to the start of another module.

44. The apparatus according to claim 43, wherein the buffer tank is configured to act as a reaction vessel.

45. The apparatus according to any one of claim 1 to 44, wherein the reagent for the capping station comprises acetic anhydride, acetyl chloride, propionic anhydride, isobutyric anhydride, phenoxyacetic anhydride, and / or / c / 7-butylphenoxyacetic anhydride.

46. The apparatus according to any one of claims 1 to 45, wherein the reagent further comprises a catalyst.

47. The apparatus according to claim 46, wherein the catalyst comprises N- methylimidazole, 4-dimethylaminopyridine, and / or pyridine.

48. The apparatus according to any one of claim 1 to 47, wherein the washing fluid comprises one or more of acetonitrile, dichloromethane, dichloroethane, chloroform, dimethoxy ethane, N,N-dimethylformamide, N,N’-dimethylacetamide, N-methyl-2- pyrrolidone, N-formylmorpholine, N-butylpyrrolidinone, cyclopentyl methyl ether, methyl tert-butyl ether, diethyl ether, tetrahydrofuran, toluene, 2-methyltetrahydrofuran, ethyl acetate, dimethyl carbonate, dioxane, methanol, ethanol, iso-propanol, water, and dimethyl sulfoxide.

49. The apparatus according to any one of claim 1 to 48, wherein the reagent delivered at the deprotection station is suitable for removing a protecting group.

50. The apparatus according to any one of claim 1 to 49, wherein the oligomer comprises an oligonucleotide.

51. The apparatus according to claim 50, wherein the reactant for the coupling station comprises a nucleoside phosphoramidite, phosphortri ester, and / or H-phosphonate.

52. The apparatus according to claim 50 or claim 51, wherein the reactant further comprises an activator.

53. The apparatus according to claim 52, wherein the activator comprises IH-tetrazole, 5- (4-nitrophenyl)-lH-tetrazole, 5-(bis-3,5-trifluoromethylphenyl)-lH-tetrazole, 5-methylthio- IH-tetrazole. 5-ethylthio- IH-tetrazole, 5-benzylthio- IH-tetrazole, 5-mercapto-tetrazole, 1- hydroxy-benzotriazole, 3-nitro-lH-l,2,4-triazole, pyridinium trifluoroacetate, imidazoliumtriflate, imidazolium perchlorate, imidazolium tetrafluoroborate, N-(methyl)imidazolium triflate. N-(phenyl)imidazolium triflate, N-(phenyl)imidazolium perchlorate, N- (phenyl)imidazolium tetrafluoroborate, N-(p-acetylphenyl)imidazolium triflate, 2- (phenyl)imidazolium triflate, 4-(methyl)imidazolium triflate, 4-(methyl)imidazolium tosylate, 4-(methyl)imidazolium trifluoroacetate, 4-(phenyl)imidazolium triflate, 4- (phenyl)imidazolium trifluoroacetate, benzimidazolium triflate. benzimidazolium tetrafluoroborate, N-(methyl)benzimidazolium triflate, 2-(phenyl)benzimidazohum triflate, 2- (phenyl)benzimidazolium perchlorate, diisopropyl ammonium tetrazolide, and / or 4,5- di cyanoimidazole.

54. The apparatus according to any one of claim 40 to 53. wherein the reagent for the oxidation station comprises iodine, tert-butyl hydroperoxide, and / or 1 O-camphorsulfonyl- oxaziridine.

55. The apparatus according to any one of claim 40 to 53, wherein the reagent for the thiolation station comprises 3-(dimethylaminomethylidene)amino-3H-l,2,4-dithiazole-3- thione, 3H-l,2-benzodithiol-3-one 1,1 -dioxide, 3-(dimethylaminomethylidene)amino-3H- l,2,4-dithiazole-3-thione, xanthane hydride, and / or N,N,N’N'-tetraethylthiuram disulfide.

56. The apparatus according to any one of claim 1 to 49, wherein the oligomer comprises a peptide.

57. The apparatus according to claim 56, wherein the reactant for the coupling station comprises an amino acid and one or more activators.

58. The apparatus according to claim 57, wherein the activator comprises benzotriazol- 1- yloxytris(dimethylamino)phosphonium hexafluorophosphate, (l-cyano-2-ethoxy-2- oxoethylidenaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate, N,N'- di cyclohexylcarbodiimide, diisopropylcarbodiimide, l-ethyl-3-(3- dimethylaminopropyl)carbodiimide, l-[bis(dimethylamino)methylene]-lH-l,2,3-triazolo[4,5- b]pyridinium 3-oxide hexafluorophosphate, O-(benzotriazol-l-yl)-N,N,N’,N’- tetramethyluronium hexafluorophosphate, l-hydroxy-7-azabenzotriazole, 1- hydroxybenzotriazole, benzotriazol-1 -yloxytripyrrolidinophosphonium hexafluorophosphate,propylphosphonic anhydride, O-(7- azabenzotriazol-l-yl)- N,N,N’,N’-tetramethyluronium tetrafluoroborate, O-(benzotriazol-l-yl)- N.N.N’.N'-tetramethyluromum tetrafluoroborate.

59. The apparatus according to any one of claim 1 to 49, wherein two or more stations act on different parts of the solid phase material concurrently.

60. A method of preparing an oligomer, comprising synthesizing the oligomer using the apparatus of any one of claims 1-59.

61. A method of quasi-continuous oligomer synthesis, comprising(a) providing a solid phase material and an apparatus for oligomer synthesis, said apparatus comprising at least one deprotection station; at least one coupling station; optionally, one or more oxidation and / or thiolation stations; optionally, one or more capping stations, and, optionally, one or more washing stations; wherein the apparatus is configured to act on one or more solid phase materials at each station to produce an oligomer; and wherein each station is at a fixed position in the apparatus and configured to deliver a reagent, a reactant, or a washing fluid comprises to a moveable solid phase material via a fluid delivery device, and(b) initiating movement of the solid phase material through the stations of the apparatus to produce an oligomer.

62. The method according to claim 61, wherein the at least one deprotection station carries out a step of deprotecting.

63. The apparatus according to claim 61 , wherein the at least one coupling station carries out a step of coupling.

64. The apparatus according to claim 61, wherein the one or more oxidation and / or thiolation stations carries out a step of oxidation or thiolation.

65. The apparatus according to claim 61, wherein the one or more capping stations carries out a step of capping.

66. The apparatus according to claim 61, wherein the one or more washing stations carries out a step of washing.

67. The method according to any one of claims 61 to 66, further comprising attaching the oligomer to the solid phase material via a linker, preferably by solid phase coupling.

68. The method according to any one of claims 61 to 67, further comprising depositing the solid phase material onto a conveyor device.

69. The method according to any one of claim 61 to 68. wherein the conveyor device is moving the solid phase material through the stations.

70. The method according to any one of claim 61 to 69, wherein deprotection comprises contacting a protected oligomer with a reagent suitable for deprotection to cause removal of the protecting group.

71. The method according to any one of claim 61 to 70, wherein coupling comprises contacting an oligomer with a reactant and optionally one or more reagents to form a linkage between the oligomer and the reactant.

72. The method according to any one of claim 61 to 71, wherein washing comprises contacting an oligomer with a washing fluid to remove residual reagent or reactant from the solid phase material.

73. The method according to any one of claim 61 to 72, further comprising delivering the reagent, the reactant, or the washing fluid to the solid phase material via a spray nozzle, a perforated nozzle, a perforated plate, or an open pipe.

74. The method according to any one of claim 61 to 73, further comprising applying a pressure gradient to the conveyor device to increase the flow of the reagent, reactant, or washing fluid.

75. The method according to claim 74, further comprising applying the pressure gradient by applying vacuum to the underside of the conveyor belt.

76. The method according to claim 74, further comprising applying the pressure gradient by applying positive pressure to the top face of the conveyor belt.

77. The method according to any one of claim 91 to 76, further comprising recovering an excess reagent, reactant, or washing fluid.

78. The method according to any one of claim 61 to 77, further comprising recovering the excess reagent, reactant, or washing fluid in a collecting tank after it has passed through the solid phase material.

79. The method according to claim 77 or claim 78, wherein the recovering comprises feeding the recovered reagent, the recovered reactant, or the recovered washing fluid back to the fluid delivery device.

80. The method according to claim 77 or claim 78, wherein the recovering comprises feeding the recovered reagent, the recovered reactant, or the recovered washing fluid to one or more additional fluid delivery devices.

81. The method according to claim 79 or claim 80, wherein the feeding to one or more additional fluid delivery devices is countercurrent feeding.

82. The method according to claim 79 or claim 80, wherein the feeding to one or more additional fluid delivery devices is cocurrent feeding.

83. The method according to any one of claim 621 to 82, further comprising releasing the oligomer from the solid support by one or more chemical cleavage reactions or by a change in pH and / or temperature.

84. The method according to any one of claim 61 to 83, further comprising monitoring an oligomer via a product analytical device configured to provide a signal indicating a solid phase material should be retained or discarded.

85. The method according to claim 84, wherein the monitoring is conducted at one or more stations.

86. The method to any one of claim 1 to 85, further comprising retaining or discarding the solid phase material based on the signal from the product analytical device.

87. The method according to any one of claim 61 to 86. further compnsing preparing multiple oligomers simultaneously.

88. The method according to any one of claim 61 to 87, further comprising collecting the one or more solid phase materials in a buffer tank at the end of a module and feeding it to the start of another module.

89. The method according to claim 88, further comprising carrying out a reaction in the buffer tank.

90. The method according to any one of claim 61 to 89, further comprising releasing the oligomer from the solid phase material.

91. The method according to any one of claim 61 to 90, further comprising depositing the one or more solid phase material onto the conveyor device.

92. The method according to any one of claim 61 to 91, wherein the oligomer comprises an oligonucleotide.

93. The method according to any one of claims 61 to 92 wherein coupling comprises contacting the nucleotide with a reactant comprising a nucleoside phosphoramidite, phosphortriester, and / or H-phosphonate.

94. The method according to claim 93, wherein the reactant further comprises an activator.

95. The method according to claim 94, wherein the activator comprises IH-tetrazole, 5- (4-nitrophenyl)-lH-tetrazole, 5-(bis-3,5-trifluoromethylphenyl)-lH-tetrazole, 5-methylthio-IH-tetrazole, 5-ethylthio-lH-tetrazole, 5-benzylthio-lH-tetrazole, 5-mercapto-tetrazole, 1- hydroxy -benzotriazole, 3-nitro-lH-l,2,4-triazole, pyridinium trifluoroacetate, imidazolium triflate, imidazolium perchlorate, imidazolium tetrafluoroborate, N-(methyl)imidazolium triflate, N-(phenyl)imidazolium triflate, N-(phenyl)imidazolium perchlorate, N- (phenyl)imidazolium tetrafluoroborate, N-(p-acetylphenyl)imidazolium triflate, 2- (phenyl)imidazolium triflate, 4-(methyl)imidazolium triflate, 4-(methyl)imidazolium tosylate, 4-(methyl)imidazolium trifluoroacetate. 4-(phenyl)imidazolium triflate, 4- (phenyl)imidazolium trifluoroacetate, benzimidazolium triflate, benzimidazolium tetrafluoroborate, N-(methyl)benzimidazolium triflate, 2-(phenyl)benzimidazolium triflate, 2- (phenyl)benzimidazolium perchlorate, diisopropylammonium tetrazolide, and / or 4.5- di cyanoimidazole.

96. The method according to any one of claims 1 to 95, wherein the step of oxidation comprises contacting an oligonucleotide with an oxidant to cause conversion of a phosphite triester to a phosphate triester.

97. The method according to any one of claims 61 to 96, wherein thiolation comprises contacting an oligonucleotide with a thiolation reagent to cause conversion of a phosphite triester to a phosphorothioate.

98. The method according to any one of claim 61 to 97, wherein capping comprises contacting an oligonucleotide with one or more reagents to cause conversion of one or more residual free hydroxy groups into esters.

99. The method according to any one of claim 96 to 98, wherein the oligomer comprises a peptide.

100. The method according to any one of claims 61 to 91, wherein the reactant comprises an ammo acid.

101. The method according to claim 100, wherein the activator comprises benzotriazol- 1- yloxy tris(dimethylamino)phosphonium hexafluorophosphate, ( 1 -cy ano-2-ethoxy-2- oxoethylidenaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate, N,N'- di cyclohexylcarbodiimide, diisopropylcarbodiimide, l-ethyl-3-(3-dimethyl aminopropyl Jcarbodilmide. l-[bis(dimethylamino)methylene]-lH-l,2,3-triazolo[4,5- b]pyridinium 3-oxide hexafluorophosphate, O-(benzotriazol- l -yl)-N.N.N'.N'- tetramethyluronium hexafluorophosphate, l-hydroxy-7-azabenzotriazole, 1- hydroxybenzotriazole, benzotriazol-1 -yloxytripyrrolidinophosphonium hexafluorophosphate, propylphosphonic anhydride, O-(7- azabenzotriazol-l-yl)- N,N,N’,N’-tetramethyluronium tetrafluoroborate, O-(benzotriazol-l-yl)- N,N,N’,N'-tetramethyluronium tetrafluoroborate.

102. The method according to any one of claim 61 to 91 or 99 to 101, wherein capping comprises contacting a peptide with one or more reagents to cause conversion of one or more residual amine into amides.

103. The method according to any one of claim 61 to 91 or 99 to 102, wherein capping comprises contacting a peptide with one or more reagents to cause conversion of one or more residual carboxylic acid groups into esters.

104. The method of any one of claims 61 to 103, wherein the method is performed continuously.

105. An oligonucleotide produced according to the method of any one of claims 61 to 98.

106. A peptide produced according to the method of any one of claims 61 to 91 or 99 to104.

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