76 results about "Diafiltration" patented technology

Provided herein are methods for making liposomal API formulations via continuous in-line diafiltration processes. Also provided herein are liposomal API formulations manufactured by the disclosed methods.

The present invention provides an improved process for isolating 2,4-bis-(2,4-dihydroxyphenyl)-6-(4-methoxyphenyl)-1,3,5-triazine (DIOPAT) from an aqueous alkaline mixture M comprising the DIOPAT, 2,4-dihydroxybenzophenone, and aluminum salts, wherein the process comprises the steps of precipitating the DIOPAT by acidifying the mixture M to a pH<1; heating the acidified mixture M to a temperature in the range of from 80° C. to 95° C.; and separating of the precipitated DIOPAT from the dissolved 2,4-dihydroxybenzophenone and the dissolved aluminum salts with a ceramic membrane by means of diafiltration.

Provided herein are methods for making liposomal API formulations via continuous in-line diafiltration processes. Also provided herein are liposomal API formulations manufactured by the disclosed methods.

Iodized micellar casein is produced by adding, to an aqueous solution of micellar casein, crystalline potassium iodide at a ratio of 1:7-1:9; holding the mixture for 5-10 minutes; adding to the mixture crystalline potassium persulphate at a ratio of 1:8-1:12 crystalline potassium persulphate to micellar casein mass; holding the mixture for 4-8 hours at a temperature of 18-25°С and a pH of 6.5-8.9 while constantly monitoring the molecular iodine content of the solution and periodically adding portions of crystalline potassium persulphate at a ratio of 1:12-1:24 crystalline potassium persulphate to micellar casein mass to continue the iodination reaction; removing residual inorganic salts from the resulting aqueous solution by microfiltration and diafiltration, followed by sterilizing microfiltration and freeze drying or spray drying. The method makes it possible to produce iodized micellar casein that is suitable for creating products for preventing iodine deficiency and optimizing iodine metabolism and for preventing and protecting against radiation poisoning.

A method of purifying a composition comprising a Group B adenovirus, for example comprising the following purification step: diafiltration of a composition comprising the Group B adenovirus using a diafiltration buffer having a conductivity of at least 180 mS cm ‑1 , for example a conductivity of 190, 200, 210, 220, 230, 240, 250, 260, 270, 280 or 290 mS cm ‑1 . Also provided are compositions obtained using the purification methods disclosed herein.

The application provides a method for extracting oil bodies by combining diafiltration and ultrafiltration, comprising the following steps: 1) soaking and crushing plant seeds containing oil bodies, separating the material residue, removing the waste residue by filtering through filter cloth, and obtaining a first mixture of protein oil bodies; 2) filtering the first mixture of protein oil bodies through a 5-kDa diafiltration membrane to obtain a second mixture of protein oil bodies; and 3) ultrafiltering the second mixture of protein oil bodies through a 5-kDa ultrafiltration membrane to obtain a third mixture of protein oil bodies. The method for extracting oil bodies by combining diafiltration and ultrafiltration has the advantages that: the method uses ultrafiltration and diafiltration technologies, the equipment has a smaller volume, energy consumption is reduced, and the method is suitable for industrial production of oil bodies. Compared with centrifugal separation products, the product obtained by the method has higher protein content, less water content and more fat content.

The present disclosure provides high viscosity ultrafiltration / diafiltration (UF / DF) and single pass tangential flow filtration (SPTFF) processes for the purification of polypeptides. A method for purifying a polypeptide from a polypeptide preparation can include the following steps in sequence: a) subjecting the polypeptide preparation to one or more purification processes and recovering a first resulting pool having a viscosity of about 5 to 40 cP; and b) subjecting the pool recovered from step a) to an SPTFF operation and recovering a second resulting pool having an operating viscosity of about 41 to 400 cP, the SPTFF operation comprising the use of membranes in either or both of a tandem membrane configuration and / or a parallel membrane configuration, where i) the operating temperature is about 15 to 55 DEG C, and ii) the feed flux is about 5 to 50 L / m2 / hr (LMH).

Large scale produced water-soluble legume proteins and obtained by: Crushing legume seeds, if necessary, degreasing the crushed legume seeds; Mixing the crushed legume seeds with water to produce a legume slurry; Adjusting the pH value of the legume slurry to a pH value between 6.8 and 7.5, preferably between 7.0 and 7.4; Separating starch and fibers by centrifugation or filtration to produce an aqueous protein solution as supernatant; Adjusting the pH value of the separated protein solution to a pH value between 7.2 and 8, Ultrafiltrating the pH-adjusted protein solution; Diafiltrating the ultrafiltration retentate with water with a pH value of 7.5-8.2 to a conductivity of the diafiltrate of no more than 30% of the conductivity of the permeate without diafiltration, i.e., to a conductivity of 1-3 mS / cm; Obtaining the diafiltered ultrafiltration protein retentate; and drying, cooling or freezing the ultrafiltration retentate, and a method for its production.

To provide, in particular, a method of producing an immunogenic composition exhibiting reduced virucidal activity, as well as the immunogenic composition and uses thereof.SOLUTION: The method in particular comprises the steps of: (a) providing a mixture with a first liquid and a recombinant protein, (b) concentrating the recombinant protein in the mixture by removing a portion of the first liquid from the mixture, and (c) processing the solution resulting from step (b) by continuous diafiltration.SELECTED DRAWING: Figure 1

The present invention discloses a system for separating trivalent antimony and pentavalent antimony using a strong cationic resin, relating to the technical field of separation of trivalent antimony and pentavalent antimony. The specific steps are as follows: Step 1: Cleaning: Using a cleaning device, clean an SCX chromatographic column with 10% by mass sulfuric acid and ultrapure water; Step 2: Diafiltration: Introducing a sample solution into the SCX chromatographic column at a flow rate of approximately 1.5 ml / min, collecting the filtrate from the SCX chromatographic column, and rinsing it with ultrapure water. The rinsing liquid and the filtrate are collected together to form a separation solution for pentavalent antimony; Step 3: Eluting the captured trivalent antimony with hydrochloric acid at a flow rate of 1.5 ml / min. The present invention involves cleaning the SCX chromatographic column with sulfuric acid and ultrapure water; diafiltration: Introducing a sample solution into the SCX chromatographic column, collecting the filtrate from the SCX chromatographic column, and rinsing it with ultrapure water. The rinsing liquid and the filtrate are collected together to form a separation solution for pentavalent antimony; and eluting the trivalent antimony with hydrochloric acid, thereby facilitating the separation of trivalent antimony from pentavalent antimony.

The present invention relates to a microporous membrane percolation detection device based on immune lateral flow, which includes an immune lateral flow channel and a microporous membrane with a pore size less than 1.2um, a solution outflow outlet is arranged at the end of the immune lateral flow channel, and the microporous membrane is attached to the solution outflow outlet; at least one labeling reagent is arranged in the immune lateral flow channel, and a capture reagent is coated on the microporous membrane. The present invention also discloses a microporous membrane percolation detection device, which includes an immune lateral flow channel and a microporous membrane with a substantially uniform pore size and a pore size greater than 20nm, a solution outflow outlet is arranged at the end of the immune lateral flow channel, and the microporous membrane is attached to the solution outflow outlet, and the microporous membrane is a hydrophilic microporous membrane; coated microspheres, a capture reagent fixed on the surface of the microspheres, and a labeling reagent are arranged in the immune lateral flow channel. Substances that are direct or indirect ligand relationships with each other are respectively arranged in the capture reagent and the labeling reagent.

The present invention relates to a method for diafiltration using a crossflow filtration unit for the continuous diafiltration of a food liquid to obtain a retentate and a permeate.

Provided herein are methods for purifying recombinant A Disintegrin-like and Metallopeptidase with Thrombospondin Type 1 Motif 13 (ADAMTS13) protein from a sample. The method comprises enriching for ADAMTS13 protein by chromatographically contacting the sample with hydroxyapatite under conditions that allow ADAMTS13 protein to appear in the eluate or supernatant from the hydroxylapatite. The methods may further comprise tandem chromatography with a mixed mode cation exchange / hydrophobic interaction resin that binds ADAMTS13 protein. Additional optional steps involve ultrafiltration / diafiltration, anion exchange chromatography, cation exchange chromatography, and viral inactivation. Also provided herein are methods for inactivating virus contaminants in protein samples, where the protein is immobilized on a support. Also provided herein are compositions of ADAMTS13 prepared according to said methods.

The present disclosure provides methods for deprotecting and purifying an oligomeric compound. The method provides for deprotection of a protecting group, which may be a trityl protecting group, and removal of a byproduct by diafiltration. Also provided are oligomeric compounds prepared by the method.

The present disclosure relates to methods for purifying proteins of interest using ultrafiltration and / or diafiltration. Specifically, the present disclosure utilizes a more accurate means for estimating the osmolality of the protein solution that has undergone an ultrafiltration and / or diafiltration operation. The method involves a mathematical model, which employs estimation of the osmolarity of the protein mixture based on the activity of water and Norrish constants for each excipient present in the protein mixture.

The invention relates to methods for extracting, purifying and concentrating mesenchymal stromal cell exosomes enriched with micro-RNA, proteins and lipids, and can be used in cosmetology and pharmacology for rehabilitation and regeneration. The technical result of the invention is a more efficient process for producing exosomes. This technical result is achieved in that the present method for extracting, purifying and concentrating exosomes from mesenchymal stromal cells includes cultivating cells, collecting the conditioned culture medium, depleting high-molecular-weight proteins, extracting, purifying and concentrating exosomes, and freezing and lyophilizing the exosome concentrate; in order to enrich the culture medium with exosomes, multipotent mesenchymal stem cells (MSCs) are grown in a medium having a lowered glucose concentration and containing platelet lysate and 5 mM of L-alanyl-L-glutamine, wherein, in the case of static cell cultivation, the growth medium is placed in a culture vessel and once a level of not less than 80% monolayer confluence is reached, the culture medium is harvested and replaced once every 24 hours, and in the case of flow-through cell cultivation, cells are placed on cell carriers in a bioreactor and 72-96 hours after the start of cultivation, the culture medium is harvested and replaced once every 24 hours, the exosome-enriched culture medium is subjected to concentration, which includes successive stages of centrifugation, tangential flow filtration and isopycnic centrifugation: in the first stage, the culture medium is centrifuged for 10 mins at 2000 g to remove large cell particles and other inclusions; in the tangential flow filtration stage, the supernatant is concentrated using hollow fibre filters having a molecular weight cutoff of 500 kDa, after which the concentrate is subjected to fivefold diafiltration with an equal volume of DPBS phosphate buffer solution using the same hollow fibre cartridge; and in the isopycnic centrifugation stage, the concentrated supernatant is mixed with a saccharose solution and deuterium oxide and subjected to isopycnic centrifugation at 100000 g and 4˚С for 75 mins, after which the concentrate is subjected to further fivefold diafiltration with a buffer solution, then the concentrate is filtered through filters with a pore diameter of 0.22 µm and 0.1 µm, the exosome concentrate is subjected to initial freezing to -75˚C in a freezing chamber, and drying is carried out in a lyophilization chamber, wherein in a first drying stage, the concentrate is cooled to -40˚С and subsequently held for 100 mins at an absolute pressure in the chamber of not more than 10 Pa, then cooled to -30˚С and subsequently held for 610 mins at an absolute pressure in the chamber of not more than 10 Pa, then cooled to -10˚С and subsequently held for 315 mins at an absolute pressure in the chamber of not more than 10 Pa, after which it is cooled to 0˚С and subsequently held for 130 mins at an absolute pressure in the chamber of not more than 10 Pa, then heated to +10˚С and subsequently held for 130 mins at an absolute pressure in the chamber of not more than 10 Pa; in a second drying stage, drying is carried out at +22˚С for not less than 130 minutes at an absolute pressure in the chamber of not more than 10 Pa.

Disclosed herein are ultrafiltration / diafiltration (UF / DF) methods comprising the use of diafiltration buffer concentrates, as well as UF / DF methods in which a composition comprising a recombinant protein is concentrated by an ultrafiltration step to a recombinant protein concentration above an intended concentration following a subsequent diafiltration step.

The disclosure describes methods for the purification of protein-enriched extracts to provide concentrates and isolates and methods for incorporation of such materials into products. The purification methods are adapted for removal of one or more of ash, metal salts, alkaloids, particulates, heavy metals, and other impurities and / or contaminants from extracts, as well as modifying the sensory characteristics (e.g., odor, color, and / or taste characteristics) of extracts. The methods generally include diafiltration, treatment with functionalized resins, and supercritical extraction. A protein composition in the form of a concentrate or isolate is provided, the protein composition including RuBisCO, F2 fraction proteins, or combination thereof extracted from a plant of the Nicotiana species, wherein the protein composition is characterized by one or more of: an ash content of less than about 15% by weight; a nicotine content of less than about 10 μg / g; and a heavy metal content of less than about 60 μg / g.

Systems, devices, and methods for managing air gaps in a semiconductor device are provided. In one aspect, a method includes removing coelomic fluid from an exterior surface of the worms using a drying agent; removing blood from the worms; performing centrifugation on the blood to generate a blood supernatant; concentrating, by a tangential flow filtration system with a single membrane, the blood supernatant to form a concentrated blood product; and performing, by the tangential flow filtration system, tangential flow filtration of a diafiltration mixture comprising the concentrated blood product.

The present invention describes a novel method for the isolation of a glycosphingolipid lacking the amide-linked fatty acyl group, the method comprising the steps of: providing a solution comprising said glycosphingolipid of formula (1) or a salt thereof, and one or more contaminants, diafiltration of said solution, thereby obtaining a diafiltration retentate (DFR) comprising said glycosphingolipid, or salt thereof, by using a membrane having a MWCO of 100-300 kDa, thereby isolating said glycosphingolipid or salt thereof from the one or more contaminants.

Provided herein are methods for analyzing the amino acid content of a therapeutic protein. In exemplary embodiments, the method comprises exchanging by diafiltration the therapeutic protein into a second solution from a first solution to obtain an exchanged protein solution; incubating the exchanged protein solution at an acidic pH at a temperature greater than 110 °C to obtain a hydrolyzed sample comprising amino acids of the therapeutic protein; combining the hydrolyzed sample with activated charcoal; and separating the hydrolyzed sample from the activated charcoal by filtration, centrifugation, or both filtration and centrifugation; and analyzing the amino acid content of the hydrolyzed sample.

To provide a fully integrated, continuous, manufacturing process for biologics manufacturing by integration of drug substance and drug product processes.SOLUTION: Provided is a method for producing a recombinant biologic therapeutic, comprising: providing a purified recombinant protein of interest; concentrating or diluting the purified recombinant protein by ultrafiltration; buffer exchanging the purified recombinant protein into a desired formulation by diafiltration; further diluting or concentrating the formulated recombinant protein by ultrafiltration until a target concentration is achieved; adding or combining at least one stability -enhancing excipient once the target concentration is achieved; subjecting the resulting bulk drug substance to filtration to reduce bioburden; subjecting the resulting bulk drug product to sterile filtration; and subjecting the sterile bulk drug product to a fill and finish operation; neither the purified recombinant protein nor the bulk drug substance being subjected to freezing and thawing unit operations.SELECTED DRAWING: Figure 1B

Disclosed herein is a purification process comprising an organic ultrafiltration / diafiltration process in which a crude mixture containing a conjugate compound to be purified is filtered through a semipermeable membrane and the filtrate from the crude mixture is replaced with an exchange medium containing an organic solvent and a buffer system.

The invention is directed to a composition of soluble placental ECM in storage solution and the methods of manufacture related thereto. The solubilized placental ECM solution is manufactured through a process of partitioning placental ECM from the placental tissue in a high acidic pH environment and performing an ultrafiltration step and diafiltration step through a tangential flow filtration system to increase the pH. The soluble placental ECM retains the native characteristics of placental ECM components and includes collagens (types I, III, V, VI, IX, and X), elastin, laminin, and fibronectin.

The present invention discloses a polycyclic polyisopentenyl phloroglucinol compound and its preparation method and application, belonging to the field of biomedicine technology. The dried aerial part of Hypericum perforatum is used as raw material, and an extract is obtained after diafiltration extraction and concentration, and the extract is suspended in water to obtain a suspension, and a petroleum ether extract is obtained after extraction and concentration; the extract is taken and subjected to positive and reverse phase silica gel column, MCI medium pressure column chromatography, Sephadex LH-20 gel column chromatography separation and high performance liquid chromatography purification to obtain a polycyclic polyisopentenyl phloroglucinol compound, and the structure and configuration of the compound are determined by means of spectral analysis technology, quantum chemical calculation and X-ray single crystal diffraction. This type of compound can inhibit TNF-α protein from exerting an immunosuppressive effect and has the use as an immunosuppressive drug.

A method for improving the harvest or purification of a target protein such as a biologic or biosimilar is provided. The method improves conventional harvest / purification methodologies by adding a chromatography step, such as a mixed-mode or ion exchange chromatography step, towards the end of the polishing phase of harvest / purification, after conventional chromatographic polishing steps such as protein A or ion exchange chromatography steps have been completed and the resultant eluate subjected to filtration, such as ultrafiltration / diafiltration. The surprising result of returning to chromatographic polishing after filtration is that all forms of high molecular weight products are reduced, facilitating the purification of target protein sufficient to meet government regulations, such as Quality Target Protein Profiles.

The present invention provides, among other things, improved methods for purifying arylsulfatase A (ASA) protein produced recombinantly for enzyme replacement therapy. The present invention is, in part, based on the surprising discovery that recombinant ASA protein can be purified from unprocessed biological materials, such as, ASA-containing cell culture medium, using a process involving as few as four chromatography columns and only one step of post-chromatographic ultrafiltration / diafiltration.

Disclosed is a high resolution affinity chromatography for analyzing a capture process and an affinity capture process using a single chromatography matrix combination, the high resolution affinity chromatography resulting in an improved resolution between closely related molecular species, and the total product yield of large-scale commercial production of heterodimer proteins (such as bispecific antibodies) is significantly improved. Furthermore, storage tank and equipment requirements are reduced by reducing salt concentration while increasing product purity and concentration without dilution, ultrafiltration or percolation capability.

The present invention provides, inter alia, improved methods for purifying arylsulfatase A (ASA) proteins that are recombinantly produced for enzyme replacement therapy. The present invention is based, in part, on the surprising discovery that recombinant ASA proteins can be purified from untreated biological materials (e.g., ASA-containing cell culture media) using processes that include only four chromatographic columns and only one step of post-chromatographic column ultrafiltration / diafiltration.

The present invention describes a novel method for the isolation of a glycosphingolipid lacking the amide-linked fatty acyl group, the method comprising the steps of: providing a solution comprising said glycosphingolipid of formula (1) or a salt thereof, and one or more contaminants, diafiltration of said solution, thereby obtaining a diafiltration retentate (DFR) comprising said glycosphingolipid, or salt thereof, by using a membrane having a MWCO of 100-300 kDa, thereby isolating said glycosphingolipid or salt thereof from the one or more contaminants.