598 results about "Anion Exchange Proteins" patented technology

The present invention provides a process for the production of erythropoietin (EPO) with high purity and with a desired profile of EPO glycol-isoforms by using a combination of specific chromatographic steps in such a manner that the starting EPO glycol-isoform profile is changed or modified. The applied chromatographic steps includes at least (a) dye affinity chromatography, and (b) hydrophobic chromatography and/or (c) anion-exchange chromatography. In a preferred embodiment, the process further includes (d) gel filtration chromatography. The present invention also provides a process for the determination of erythropoietin (EPO) glycol-isoform profile in an EPO containing composition.

A method is disclosed for separating a polypeptide monomer from a mixture comprising dimers and/or multimers. The method comprises applying the mixture to either a cation-exchange chromatography resin or an anion-exchange chromatography resin and eluting the mixture at a gradient of about 0-1 M of an elution salt, wherein the monomer is separated from the dimers and/or multimers present in the mixture.

The invention relates to bone health compositions comprising an acidic protein fraction of milk, to a method of producing said bone health composition, to methods of treatment comprising said bone health compositions and to medicinal uses of said bone health compositions. One broad aspect of the invention provides a bone health composition comprising an acidic protein fraction derived from milk, from a component of milk, from whey, from hydrolysates thereof, or from a combination thereof, or from a combination thereof wherein the composition does not comprise caseinoglycomacropeptide (CGMP). Another broad aspect provides a method of manufacturing the composition of the invention using anion exchange chromatography.

The current invention provides methods (e.g., large-scale processes) for the production of nucleotide sugars, which are modified with a polymeric modifying group, such as poly(alkylene oxide) moieties (e.g., poly(ethylene glycol) or poly(propylene glycol)) moieties. A typical process of the invention includes anion exchange chromatography followed by an ultrafiltration procedure, such as tangential flow filtration. The process of the invention provides modified nucleotide sugars in unexpectedly high purity and high overall yields.

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.

An improved process for the purification of antibodies from human plasma or other sources is disclosed. The process involves suspension of the antibodies at pH 3.8 to 4.5 followed by addition of caprylic acid and a pH shift to pH 5.0 to 5.2. A precipitate of contaminating proteins, lipids and caprylate forms and is removed, while the majority of the antibodies remain in solution. Sodium caprylate is again added to a final concentration of not less than about 15 mM. This solution is incubated for 1 hour at 25° C. to effect viral inactivation. A precipitate (mainly caprylate) is removed and the clear solution is diluted with purified water to reduce ionic strength. Anion exchange chromatography using two different resins is utilized to obtain an exceptionally pure IgG with subclass distribution similar to the starting distribution. The method maximizes yield and produces a gamma globulin with greater than 99% purity. The resin columns used to obtain a high yield of IgG retain IgM and IgA. IgA and IgM may be eluted from these resins in high yield and purity.

A novel strain of lactic acid bacteria was found to be heat resistant and able to grow in a sulfur-limiting medium (SLM) containing a high concentration of sodium selenite. The microorganism is a non-spore forming and Gram-positive coccus, which is identified with >90% confidence using the API biochemical and sugar fermentation tests, ribotyoing and 16S rRNA sequencing as Pediococcus pentosaceus SP80. In the current study, P. pentosaceus SP80 grown on SLM containing 250 ppm sodium selenite produced both organic and inorganic forms of selenium. These selenium compounds can be separated using an anion exchange chromatography technique. The concentrations of selenium detected in the organic and inorganic fractions were 4.34 and 21.7 ppm, respectively. Selenium-enriched bacteria are useful as a source of selenium for supplementing the diets of animals and humans. Animals fed efficacious amounts of the selenium-enriched bacteria show improved feed conversion rates and higher levels of glutathione peroxidase (GPx) activity in heart, kidney and liver tissues indicating an increased absorption and retention of selenium over control diets.

The invention belongs to the biochemistry technical field, concretely relates to a method for purifying anti-HER2 or/and anti-HER3 antibody proteins. The method of the invention comprises the following the steps: clarifying a broth, carrying out affinity chromatography, carrying out inactivation on virus by acidic pH value, carrying out cation exchange chromatography, carrying out anion exchange chromatography; and filtering virus; wherein, the step of virus inactivation by acidic pH value and the step of virus filtration can be inserted at any position of the step after the affinity chromatography. The method of the invention comprises the following advantages that: 1) the chromatography step can be reduced to three steps, thereby the production efficiency can be enhanced and the production cost can be minimized. 2) the step of virus filtration is added, thereby the security can be raised.

Purification of α-1 proteinase inhibitor (α-1 PI) from solutions comprising α-1 PI is accomplished using hydrophobic interaction chromatography (HIC). In some embodiments, purification of α-1 PI is accomplished by precipitation of contaminating proteins from a starting solution comprising α-1 PI, such as human plasma, followed by anion exchange resin chromatography prior to HIC. Further purification may be accomplished by an optional cation exchange chromatography subsequent to anion exchange chromatography but prior to HIC. Some embodiments of the invention also include virus removal and/or inactivation by methods such as nano filtration and such as contact with a non-ionic detergent. The methods of the present invention result in greater yield, purity, and pathogenic clearance of plasma fractions than known methods.

The present invention describes a method for separating or partially separating heteroduplex and homoduplex DNA molecules in a mixture. In the method, the mixture is applied to an anion-exchange chromatography medium. The heteroduplex and homoduplex molecules are eluted with a mobile phase containing an eluting salt, including an anion and a cation, a buffer, and preferably including an organic solvent. The eluting is carried out under conditions effective to at least partially denature the heteroduplexes (e.g., thermal or chemical denaturing) resulting in the separation of the heteroduplexes from the homoduplexes. The method has many applications including, but not limited to, detecting mutations and comparative DNA sequencing.

The present invention relates to a method for purifying recombinant human serum albumin (rHSA) protein. The method comprises the following steps: fermented liquid containing rHSA is processed by a ceramic membrane, supernatant liquid is orderly purified by high salt cation exchange chromatography, hydrophobic layer exchange chromatography and weak anion exchange chromatography, and purified rHSA is obtained. The present invention is characterized in that solution processed by high salt cation exchange chromatography is processed by borate and then filtered by hollow fibers. The rHSA obtained can be used for producing vaccines for humans against viruses with a cell culture method, particularly rabies vaccines.

The invention discloses triple-helical Tremellan, a preparation method and application thereof. The preparation method comprises the following steps of: extracting Tremellan components through water extraction and alcohol precipitation, removing protein by using an enzyme-Sevage combined method, performing dialysis, and purifying by using anionexchange chromatography and gel filtration chromatography, and performing vacuum freeze drying, separation and purification on the components to obtain the Tremellan. By the method for preparing the Tremellan, the natural structure and activity of the Tremellan are not influenced; and the method has low requirement on equipment and low cost, and is suitable for large-scale promotion, development and use in industrial production. The purified Tremellan is subjected to component analysis, structure identification and antioxidant activity research, and results show that the Tremellan has high activity of removing hydroxyl radicals and is a potential antioxidant substance.

The invention relates to a method for preparing bonito stick protein hydrolysate with an effect of reducing uric acid. The method comprises the following main steps of raw material heat treatment, restriction digestion, membrane separation-anion exchange chromatography-gel filtration chromatography separation, concentration and spray drying so as to obtain the bonito stick protein hydrolysate with an effect of reducing uric acid. The amino acid analysis indicates that the zymolyte peptide fragment primary amino acid sequence contains four amino acids, namely histidine, arginine, lysine and threonine, the total mass content of the four amino acids is 70 percent of the total amino acid content of zymolyte. MALDI-TOF-MS mass spectrum determines that the molecular weight of the main peptide effective ingredient is less than 700Da. In-vitro uric acid reduction experiments prove that the bonito stick protein hydrolysate has a remarkable effect of inhibiting generation of uric acid, and has an inhibition rate over 50 percent; an oteracil potassium molded hyperuricemic rat animal model indicates that the bonito stick protein hydrolysate can be used for remarkably reducing the level of serum uric acid and serum creatinine of rats, and shows a relatively good kidney protecting effect.

The invention provides a preparation method of Sublancin antibacterial peptide, which comprises the following steps: (1) inoculating bacillus subtilis into a medium for fermentation, filtering the fermentation liquor; (2) orderly performing centrifugation, cation exchange chromatography, hydrophobic chromatography, size exclusion chromatography, and anion exchange chromatography of the fermentation liquor in step (1), finally performing ultrafiltration concentration and drying to obtain the Sublancin antibacterial peptide. The method of the invention is low in cost, increased in production efficiency, and high in product purity; the prepared Sublancin antibacterial peptide has purity of up to more than 99%; the process is simple, and easy to popularize and apply, and can realize large-scale production.

The invention relates to a process for the purification of an Fc-fusion protein having a pI between 6.9 and 9.5 comprising protein A or G affinity chromatography, cation exchange chromatography, anion exchange chromatography and hydroxyapatite chromatography.

The present invention discloses a preparation method of bloodsucker extract, in which the bloodsucker is added with water and homogenized, and then the bloodsucker extract is obtained by controllable enzymolysis, organic solvent precipitation, ultrafiltration and anion-exchange chromatography. The bloodsucker extract can be adopted to prepare a medicine with anticoagulation, thrombolytic ability, inhibition on platelet aggregation and neuroprotection effect. The present invention, according to the modern technological requirements of Chinese medicine, adopts the technology of controllable enzymolysis for extracting the peptide component, the separating technology of ultrafiltration and gel chromatography for purification further, and the biological pharmacy technology, thus obtaining the extract with higher pureness and stronger activity than the traditional bloodsucker extract. The method breaks through the bottleneck of the traditional animal medicine in extraction and separation, solves the problem of the traditional animal medicine that the amount of raw powder is large and the taste is fishy, improves the activity and quality controllability of the animal medicine extract further and greatly improves the utilization rate of medicinal materials.

The invention discloses a preparation method of recombinant human insulin, which relates to the protein polypeptide type medicine field. The technical problem needed to be solved by the invention aims at providing a preparation method for Escherichia coli to express the recombinant human insulin in order to overcome the technical disadvantages in the prior art of using the poisonous and harmful substance CNBr and having a complex production process. The preparation method for the recombinant human insulin of the invention comprises the steps of: thallus crashing, inclusion body dissolving, proinsulin sulfonating, purifying proinsulin, and proinsulin renaturation, enzyme cutting transmission, etc., and the preparation method is characterized in that during the inclusion body dissolving process, the -SH of proinsulin is transformed into -SSO3<->, and then human insulin is obtained through purifying the sulfonated proinsulin, reduction renaturation and enzyme cutting transmission by positive ion, and then through the negative ion exchange layer purification. The preparation method of the recombinant human insulin of the invention does not use the CNBr and has simple technique with high human insulin yield coefficient and purity.

Process for preparing a purified immunoglobulin preparation. The process comprises the steps of subjecting a crude immunoglobulin solution to caprylic acid treatment, removing protein aggregates and viruses from the immunoglobulin solution, subjecting the immunoglobulin solution to anion exchange chromatography in order to purify the immunoglobulin, filtering the immunoglobulin solution thus obtained on a virus-removal filter to produce an eluate containing immunoglobulin, and recovering the immunoglobulin. By combining caprylic acid treatment and precipitation with a protein precipitant the level of aggregated proteins and viruses is effectively reduced and a truly virus safe preparation is provided after filtration.

The invention provides a method for extracting rabies virus, and is to solve the defects that great discrepancy of quality indices of different batches occurs; removal of remaining DNA becomes difficult; the protein content of the remaining host is too high; and great side effects appear clinically when the single method of molecular sieve gel chromatography is adopted for extracting rabies virus vaccine. The essential of the invention is that a hollow fiber ultrafiltration column or ultrafiltration membrane with the molecular weight cut-off of 750KD or 500KD is used to condense and partially purify harvested liquid of virus; anion exchange chromatography or molecular sieve gel chromatography is adopted to separate and purify samples; molecular sieve gel chromatography or anion exchange chromatography is adopted to separate and purify samples got in step (2). The method for extracting rabies virus has the characteristics of great productive capacity, high product quality, excellent batch stability, being remarkably effective in removal of remaining DNA and HCP, and reducing the potential safety hazard of vaccine.

This invention provides a process for the continuous alkaline lysis of a bacterial suspension in order to harvest pDNA. It further provides for optional additional purification steps, including lysate filtration, anion exchange chromatography, triplex affinity chromatogragphy, and hydrophobic interaction chromatography. These optional purification steps can be combined with the continuous lysis in order to produce a highly purified pDNA product substantially free of gDNA, RNA, protein, endotoxin, and other contaminants.