Abstract: Contaminant viruses can be efficiently removed almost without losing the activity of protein by subjecting a plasma protein composition having a high risk of viral contamination to a treatment with a porous membrane having a pore size greater than a single-particle size of the virus, particularly by subjecting a plasma protein composition to a fractionation treatment by precipitation, before the porous membrane treatment. Particularly, a fibrinogen composition substantially free of non-enveloped viruses, Parvovirus among others, can be provided. By the application of the present invention, a safe plasma protein preparation free of viruses can be conveniently provided.

Abstract: A preparative method of isolating a preselected whey protein or group of whey proteins from a solution is provided. The method comprises the following steps: (a) contacting a whey protein solution with the preselected ion exchanger for a time and at a temperature sufficient to enable the preselected whey protein to be adsorbed; wherein the whey protein solution has (1) a protein content in the range of about 5% to about 20% by weight, (2) a pH of a preselected level, which is the level at which the preselected whey protein or group of whey proteins selectively binds to the preselected ion exchanger, and (3) a reduced ionic strength; and (b) recovering either or both of the following: (1) the whey protein component adsorbed in step (a), and (2) the breakthrough whey protein component not adsorbed in step (a).

Abstract: A purification process for large-scale production of Gc-globulin is described. The source of Gc-globulin is preferably a crude plasma fraction but can be any solution, suspension or supernatant containing Gc-globulin, e.g., a milk product, colostrum or a fermentation broth. The Gc-globulin can be plasma-derived or produced by a genetic modified organism. The process includes two key elements: purification by series of ion exchange chromatography steps, and performing at least two virus-reduction steps. A diagnostic method to measure the free Gc-globulin in a patient blood sample, a use of Gc-globulin in medicine and the preparation of a Gc-globulin medicinal product is also provided. The product can be used in therapy for patients with circulatory disorders and complications, i.e., where it is contemplated that said patients would benefit from the administration of Gc-globulin.

Abstract: A process is disclosed for the preparation of an essentially tetramer-free, substantially stroma-free, polymerized, pyridoxylated hemoglobin. Also disclosed is an essentially tetramer-free, substantially stroma-free, polymerized, pyridoxylated hemoglobin product capable of being infused into human patients in an amount of up to about 5 liters.

Abstract: Filtration methods comprise virus-filtering a solution containing a least one macromolecule. In one embodiment, the total salt content of the solution is within the range of from about 0.2 M to 2 M. In another embodiment, the total salt content of the solution is within the range of from about 0.2 M up to saturation with a salt, and the salt is selected from the group consisting of sodium chloride, potassium chloride, sodium acetate, sodium citrate, sodium phosphate, potassium dyhydrophosphate and combinations thereof. In yet another embodiment, the total salt content of the solution is within the range of from about 0.2 M up to saturation with the salt and the macromolecule is in solution during the virus filtering.

Abstract: A method of extracting and purifying recombinant protein(s) from transgenic sugarcane is disclosed. Fractioning of sugarcane juice that has been extracted from the cane stalks is obtained by using a multiple stage filtering process that uses multiple stages of decreasing porosity (preferably screening) followed by preferably membrane type filters, ion exchange, membrane adsorber, and chromatographic processes.

Abstract: A continuous method for preparing proteosome-amphiphilic determinant vaccines for parenteral or mucosal administration using diafiltration or ultrafiltration technology. The amphiphilic determinants include lipopolysaccharides from gram negative bacteria, e.g. S. flexneri, P. shigelloides and S. sonnei. Proteosomes are obtained from group B type 2b meningococci. The active proteosome-amphiphilic determinant complexes (non-covalent complexes) of the vaccine are formed using diafiltration or ultrafiltration to remove the detergent under non-static conditions. The use of diafiltration or ultrafiltration decreases processing time and the opportunity for contamination and further permits the use of ambient temperature and efficient scale-up. In addition, the process permits the reliable and continuous monitoring of the dializate which enhances the efficiency of the entire process.

Abstract: The invention concerns a method for elimination of viruses from a biological preparation wherein initially enveloped viruses are eliminated by a solvent-detergent step, then the solvent-detergents are removed by a resin composed of silicon beads and finally the preparation is nanofiltered.

Abstract: Improved process for preparing a kappa-caseino glycomacropeptide which comprises adjusting the pH of a solution of a milk starting material to below 4, cold ultrafiltration and concentration of the starting material on a spiral filter. This process is usable in industrial scale without fouling problems and without damaging the usable milk material by-product.

Abstract: A method is provided for removing citrate, aluminum, and other multivalent ions and contaminants from proteins by adjusting the pH of a solution containing the protein to a pH from about 7 to about 10, and diafiltering the pH-adjusted solution against aqueous solutions which have a low level of ions.

Abstract: Pertactin (formerly 69 kDa protein) is recovered in stable biologically pure form having no detectable adenylate cyclase activity from fermentation broth from the fermentation of Bordetella pertussis as well as from the cells. The broth is processed to selectively remove pertussis toxin (PT) and filamentous haemagglutinin (FHA), the pertactin is precipitated by ammonium sulphate and the precipitate is dissolved in buffer at pH 6.0 to 8.5, the solution then is passed through hydroxyapatite and ion-exchange chromatograph columns before final ultrafiltration. Cells are extracted with urea and the extract ultrafiltered and diafiltered. The pertactin is precipitated from the extract and the precipitate processed as above. In a variation, the broth is contacted with ammonium sulphate to precipitate pertactin, PT and FHA, the precipitate is dissolved and the PT and FHA selectively removed, before the solution is passed to the chromatograph columns.

Abstract: An isolated and purified outer membrane protein of a Moraxella strain, particularly M. catarrhalis, has a molecular mass of about 200 kDa. The about 200 kDa outer membrane protein as well as nucleic acid molecules encoding the same are useful in diagnostic applications and immunogenic compositions, particularly for in vivo administration to a host to confer protection against disease caused by a bacterial pathogen that produces the about 200 kDa outer membrane protein or produces a protein capable of inducing antibodies in a host specifically reactive with the about 200 kDa outer membrane protein.

Abstract: The invention is a corn product removal process that successfully extracts oil and zein from dry-milled corn. Oils and zein are extracted from corn using ethanol. Corn solids are separated from the ethanol, oil and zein mixture produced in the step of extracting. Thereafter, the ethanol, oil and zein mixture are membrane filtered to restrain zein from the mixture and pass an oil and ethanol mixture. At least one of zein or oil is then selected to be separated for an output corn product.

Abstract: The present invention relates generally to a method of separating one or more components from a protein mixture. More particularly, this invention is directed to a method of separating one or more components of blood plasma comprising one or more filtration steps using a cellulose-based filter aid. The present invention is useful in the preparation of therapeutics, in particular plasma-based therapeutics for use in humans.

Abstract: An antigenic composition is derived from surface extracted protein of cell wall surfaces and the culture supernatant extract of Staphylococci. The strain is highly virulent and &bgr; hemolytic on blood agar plate. They are mixed and purified by ion exchange and gel filtration column chromatography. This preparation method can be used in all Gram-positive bacilli. The antigens, having molecular weight of about 10,000-70,000 are certain kinds of glycoprotein comprising proteins (ca. 10-20%) and carbohydrates (ca. 75-90%). They are extracted from the cells with the use of a hypertonic buffer solution of pH 6.5-8.5 at a temperature below the denaturing point of the antigenic protein, and are salted out with ammonium sulfate (ca. 65-85%). The culture supernatant is extracted in the same way by salting out with ammonium sulfate (ca. 65-85%). The antigen fraction from both cell wall and culture supernatant is obtained in such an operation. The antigen can be used as a preparation antigen to Staphylococcus antigen.

Abstract: The invention relates to a method of removing endotoxin from preparations of alpha-1-acid glycoprotein (orosomucoid) by contact with a finely divided non-toxic resin such as fumed silica. The invention also relates to a purification process for alpha-1-acid glycoprotein which includes this deprogenation step, and to the depyrogenated product and its clinical uses.

Abstract: Endotoxin binding/neutralizing proteins capable of binding endotoxin in vivo, thereby neutralizing the toxic effect or bioactivity of endotoxin which are isolated from a horseshoe crab such as Limulus polyphemus, pharmaceutical compositions and pharmaceutical uses of the proteins, a method of purifying the proteins and an assay for endotoxin based on the proteins, are disclosed.

Abstract: The present invention relates to cartilage extracts and to a method of producing the same. Shark cartilage extracts having anti-angiogenic, anti-tumor, anti-inflammatory and anti-collagenolytic activities have been obtained by an improved process. The process comprises the steps of obtaining a crude cartilage extract in an aqueous solution, this crude extract being fractionated to recover molecules of a molecular weight less than about 500 kDa. Some of the biologically active components of the extract are prepared by further fractionation. The cartilage extract can be used for treating diseases or conditions having etiological components selected from the group consisting of tumor proliferation, angiogenesis, inflammation, metalloprotease activity and collagenolysis. Several cosmetic applications based on the capacity of the liquid extract to improve skin conditions are also disclosed. A simple and efficient process for the preparation of cartilage extracts is also disclosed.

Abstract: A system is described for conjugating, isolating, and purifying proteins. The system contains an ultrafiltration apparatus which is connected with a reaction vessel reservoir.

Abstract: A process is disclosed for the preparation of an essentially tetramerfree, substantially stromafree, polymerized, pyridoxylated hemoglobin. Also disclosed is an essentially tetramerfree, substantially stromafree, polymerized, pyridoxylated hemoglobin product capable of being infused into human patients in an amount of up to about 5 liters.

Abstract: This invention relates to methods for isolating highly-purified mixtures of natural type I interferons from white blood cells. The invention also relates to highly-purified mixtures of natural type I interferons which resemble natural type I interferon in that it includes 9 subtypes, i.e., alpha-1, alpha-2, alpha-5, alpha-7, alpha-8, alpha-10, alpha-14, alpha-21 and omega, giving rise to possibly 20 molecular species, including alpha-1a, alpha-1new, alpha-2a, alpha-2b, alpha-2c, alpha-5, alpha-5LG, alpha-7, alpha-8a, alpha-8c, alpha-10a, alpha-14a, alpha14-b, alpha 14-c, alpha-14LG, alpha-21a, alpha-21b, alpha-21c, omega and omega LG.

Abstract: The invention herein provides a method for recovering a polypeptide comprising exposing a composition comprising a polypeptide to a reagent which binds to, or modifies, the polypeptide, wherein the reagent is immobilized on a solid phase; and then passing the composition through a filter bearing a charge which is opposite to the charge of the reagent in the composition, so as to remove leached reagent from the composition.

Abstract: The present invention relates to the C3b/C4b receptor (CR1) gene and its encoded protein. The invention also relates to CR1 nucleic acid sequences and fragments thereof comprising 70 nucleotides and their encoded peptides or proteins comprising 24 amino acids. The invention further provides for the expression of the CR1 protein and fragments thereof. The genes and proteins of the invention have uses in diagnosis and therapy of disorders involving complement activity, and various immune system or inflammatory disorders. In specific embodiments of the present invention detailed in the examples sections infra, the cloning, nucleotide sequence, and deduced amino acid sequence of a full-length CR1 cDNA and fragments thereof are described. The expression of the CR1 protein and fragments thereof is also described. Also described is the expression of a secreted CR1 molecule lacking a transmembrane region.

Abstract: A high quality soy protein concentrate (SPC) was produced by a process of enzyme treatment combined with ultrafiltration. Soy flour, the starting material, was enzymatically treated with commercial pectinases and diafiltered with a porous stainless steel ultrafiltration system. The resulting product had reduced levels of physic acid and nucleic acids due to contaminant phytase and nuclease activity in the pectinase enzymes. The functionality of the SPC was improved due to increased solubility compared to conventional soy isolates produced by acid precipitation. High performance liquid chromatography gel filtration profiles indicated that the proteins in the SPC remained intact. The SPC also had reduced flavor when compared to the original soy flour according to gas chromatography flavor profiles and sensory evaluation.

Abstract: A process produces a water-soluble, naturally folded eukaryotic polypeptide containing two or several cysteines linked by disulfide bridges. This process involves culturing prokaryotic cells, a) in which the prokaryotic cells contain an expression vector which encodes the polypeptide which contains a prokaryotic signal sequence at the N-terminus, b) under conditions under which the polypeptide is secreted into the periplasm or the medium, c) cleaving the signal sequence and isolating the polypeptide from the periplasm or the medium. In this process, the culturing is carried out in the presence of arginine or a compound of the formula I R2—CO—NR1 (I) in which R and R1 represent hydrogen or a saturated or unsaturated branched or unbranched C1-C4 alkyl chain and R2 represents hydrogen, NHR1 or a saturated or unsaturated branched or unbranched C1-C3 alkyl chain, is suitable for the recombinant production of polypeptides in prokaryotes in a high yield.

Abstract: The present invention relates to a process for purifying immunoglobulin G from a crude immunoglobulin-containing plasma protein fraction. Said process includes a number of steps of which the anion exchange chromatography and the cation exchange chromatography are preferably connected in series. An acetate buffer having a pH of about 5.0-6.0 and having a molarity of about 5-25 mM is preferably used throughout the purification process. The invention further comprises an immunoglobulin product which is obtainable by this process. The invention also relates to an immunoglobulin product which has a purity of more than 98%, has a content of IgG monomers and dimers of more than 98.5%, has a content of IgA less than 4 mg of IgA/l, and contains less than 0.5% polymers and aggregates. Said product does not comprise detergent, PEG or albumin as a stabilizer. The product is stable, virus-safe, liquid and ready for instant intravenous administration.

Abstract: A method of separating a soluble milk component from milk is disclosed. The method involves the use of tangential flow filtration across a membrane to form a retentate and a permeate, combining the permeate with the original milk sample, and repeating this procedure until the milk has been sufficiently purified. Preferably, the milk is combined with a chelating agent, such as EDTA, to improve the purification efficiency. This procedure is advantageously employed with milk from transgenic animals which have been genetically altered to express exogenous proteins, such as therapeutic proteins, in their milk.

Abstract: Concentrated monoclonal antibody preparations for administration to humans are described in which the antibody is present at a concentration of greater than 100 mg/ml and as high as 350 mg/ml.

Abstract: A process for preparing a protein-polysaccharide conjugate includes reacting a protein with a polysaccharide to produce a mixture including a protein-polysaccharide conjugate and free protein. At least one unreacted reagent or low molecular weight component is removed from this mixture, without removing all of the free protein, to provide a purified mixture that contains the protein-polysaccharide conjugate and free protein. This purified mixture can be used as a conjugate vaccine, immunogen, or immunological reagent. Keeping the free protein in the purified mixture with the conjugate saves time and money in the conjugate production process. In another aspect of the invention, the purified mixture of the protein-polysaccharide conjugate and free protein is reacted with a hapten to produce a conjugate mixture including a hapten-protein conjugate and a hapten-protein-polysaccharide conjugate.

Abstract: In a method of purifying whey separated from lactic acid fermentation liquid by electrodialysis wherein said whey contains angiotensin-converting enzyme inhibiting peptides, the improvement which comprises using an anion exchange membrane having a permeability of diffusion coefficient in the range of 3.0 to 9.0×10−6 cm/sec. An anion exchange membrane having a permeability of diffusion coefficient in the range of 5.0 to 7.0×10−6 cm/sec. is more efficiently used. The product is particularly suited to produce granules and tablets.

Abstract: The present invention relates to stable compositions useful as primary standards and calibrators and controls comprising a cardiac troponin I (cTnI) such as native, recombinant, addition and deletion forms thereof, whether or not complexed with other troponin subunits such as TnC and/or TnT, in an inactivated human serum. The compositions are obtained by incubating troponin complexes with human serum. The compositions are characterized by an immunodetectability ratio of epitopes on the N-terminal segment to epitopes on the C-terminal segment substantially equivalent to that of pooled, fresh serum from acute myocardial infarction patients.

Abstract: This invention relates to a microporous or macroporous affinity filtration membrane wherein the matrix of the membrane is composed of chitin and the pores are made by dissolution of porogen during the preparation of the membrane. The invention also relates to a method for the purification of wheat germ agglutinin using chitin microprous or macroporous membranes.

Abstract: A method for producing an immunoglobulin preparation for intravenous injection, which comprises the steps of: fractionating an immunoglobulin-containing solution with 4 to 10 w/v % of polyethylene glycol having a molecular weight of from 1,000 to 10,000, at a pH value of from 4.5 to 6.5, an ionic strength of from 0.0001 to 0.1 M and a temperature of from 0 to 4.degree. C. to recover a supernatant fraction; and concentrating the supernatant fraction at a pH of from 3.5 to 5.0.

Abstract: A process and apparatus for purifying one or more target substances from a source liquid, employing one or more cross-flow filter elements, and one or more types of chromatography resins, in combination, to provide purification with advantageous yield, product purity, and cost -and time-efficiency.

Abstract: The present invention aims at providing a method for preparing dermonecrotic toxin, an improved toxoid of dermonecrotic toxin produced by Bordetelia and a toxoid mixture comprising said improved toxoid of dermonecrotic toxin produced by Bordetella and a toxoid of dermonecrotic toxin produced by Pasteurella. There is provided a method for partially purifying dermonecrotic toxin which comprises bringing a dermonecrotic toxin-containing solution into contact with a chromatographic gel sulfated by direct sulfation or a chromatographic gel to which a sulfated molecule is covalently bonded to thereby make the dermonecrotic toxin adsorbed on the gel, and eluting the adsorbed dermonecrotic toxin from the gel.

Abstract: An immunoglobulin preparation comprising an immunoglobulin, wherein the preparation contains, as a contaminant, serum albumin in an amount not greater than 10 .mu.g per 50 mg of immunoglobulin; and/or wherein the preparation is free of fine particles which can serve as a nucleus of insoluble foreign matter, and processes for preparing the immunoglobulin preparation are described.

Abstract: An isolated and purified non-denatured outer membrane protein CD which is that of or corresponds to that isolatable from a Branhamella strain, particularly B. catarrhalis, is isolated from a bacterial strain by fractionating a cell lysate formed by disrupting a cell mass of the bacterial strain by centrifugation to provide a pellet and a discard supernatant containing a large proportion of soluble bacterial proteins. The pellet is selectively extracted to remove the remaining soluble proteins, the membrane proteins other than CD and other contaminants such as lipopolysaccharide and phospholipids. The remaining CD-containing pellet is dispersed and solubilized and then fractionated by centrifugation to remove the remaining cell debris.

Abstract: The invention herein provides a method for recovering a polypeptide comprising exposing a composition comprising a polypeptide to a reagent which binds to, or modifies, the polypeptide, wherein the reagent is immobilized on a solid phase; and then passing the composition through a filter bearing a charge which is opposite to the charge of the reagent in the composition, so as to remove leached reagent from the composition.

Abstract: A process is described for the purification of recombinant interleukin-8, which allows the production of interleukin-8 under Good Manufacturing Practice (GMP) conditions.

Abstract: The invention relates to synthetic calibrators for immunological tests, analyte-specific epitopes being coupled to other proteins or to synthetic carriers.

Abstract: A peptide such as in a cell-free fermentation medium is ultrafiltered with a membrane having a molecular weight cut-off of at least approximately two times greater than the molecular weight of the peptide. The peptide is retained by the membrane, and is desalinated and concentrated to provide prepurification of the peptide. A preferred temperature for ultrafiltration is about 5 to about 15.degree. C. Permeate flow rate may be about 10 l/m.sup.2 /h to about 35 l/m.sup.2 /h, and permeate conductivity may be less than about 10 mS/cm. The permeate may be recycled until peptide concentration is constant. A cell-free fermentation medium containing hirudin from a recombinant microorganism such as Saccharomyces cerevisiae is ultrafiltered with a membrane having a molecular weight cut-off of about 20 kD to about 30 kD.

Abstract: The present invention is related to the separation of whey proteins, particularly the sequential separation of whey proteins into separate fractions through the use of chromatography. The present invention further provides methods and compositions for the sequential separation of whey proteins, as well as their use in various products. The present invention also provides methods and compositions for the cleaning of chromatography resins used in the separation of whey proteins.

Abstract: Methods for producing immunoglobulins and in particular anti-D immunoglobulin substantially free of virus and product resulting therefrom. Specifically provided are methods for nanofiltration of the anti-D immunoglobulin in high ionic strength buffer and with excipient such as polysorbate 80. Additional steps include diafiltration to concentrate the anti-D protein and reduce the concentration of excipient present.

Abstract: The present invention relates to a method for industrial production of a peptide preparation having specific specifications by hydrolysis of a protein material, preferably based on whey. The method comprises several steps, which makes it easy to control the method so as to obtain a product which, e.g. because of low mineral content, is well suited for peritoneal dialysis and parenteral feeding. The method gives a high yield.

Abstract: To provide a method of efficiently separate protective components of Bordetella pertussis. On the basis of differences in adsorbability to calcium phosphate gel formed by adding calcium ions to a Bordetella pertussis culture in the presence of excess phosphate ions, protective components of Bordetella pertussis are separated from the Bordetella pertussis culture. Traditionally, protective components of Bordetella pertussis have been separated using different purification methods for the respective components. According to the present invention, the use of the same means of purification for all subject components makes it possible to purify each component with high efficiency and high recovery rate, an aspect very advantageous for industrial production. It is also possible to efficiently produce an improved purified pertussis component vaccine comprising an effective combination of pertussis filamentous hemagglutinin (FHA), pertactin (PRN, 69K-OMP), pertussis fimbriae (FIM) and pertussis toxin (PT).

Abstract: A process is provided for the preparation of albumin which has extremely low levels of or is essentially free of colorants, metal ions, human proteins, fragments of albumin, polymers or aggregates of albumin, and viruses, and which is relatively non-glycated, relatively high in free thiol and with an intact C-terminus. The process comprises passing albumin (preferably expressed and secreted by transformed yeast) through two chromatography purifications, ultrafiltering the product, passing through two further chromatography steps and again ultrafiltering the product.

Abstract: The present invention relates to cartilage extracts and to a method of producing the same. Shark cartilage extracts having anti-angiogenic, direct anti-tumor proliferating, anti-inflammatory and anti-collagenolytic activities have been obtained by an improved process. The process comprises the steps of obtaining a homogenate of cartilage in an aqueous solution, this homogenate being centrifuged and further fractionated to obtain a total extract having molecules of a molecular weight comprised between 0 to 500 KDa. The composition of the liquid extract has then been investigated by different ways. Further fractionation of this extract led to the preliminary characterization of some of its active components. Due to the multiplicity of biological activities of the total liquid extract, it can be used for treating numerous diseases or conditions such as those having components selected from the group consisting of tumor proliferation, angiogenesis, inflammation and collagenolysis.

Abstract: Described is a process for making a liposomal, ion-exchange whey protein and products thereof, which result in the sustained release of amino acids into the body's circulation to generally promote skeletal muscle protein synthesis, decrease body fat in association with diet modification and improve exercise performance. The whey protein is preferably encapsulated in a liposome using a cold, or non-heated, process. After the liposomal, ion-exchange whey protein has been prepared, it is then preferably lyophilized to deliver macronutrients for use as a sports nutrition supplement and for use in medical or clinical catabolic applications.

Abstract: Methods are herein provided for the isolation and purification of zeamatin, an antifungal protein from corn. The subject methods use capture chromatography and reverse phase chromatography. The methods herein described is superior to prior art techniques as it the eliminates ammonium sulfate precipitation and centrifugation steps.

Abstract: A method is provided for cleansing a protein from multivalent metal ions bound thereto, these ions being released from the protein by exchanging the ions with monovalent metal ions, whereafter the multivalent metal ions are removed. The release and removal of these ions is effected, in particular, by diafiltration or gel filtration processes.