Abstract: Methods are provided for improved production of hIL-3 either in glycosylated form from mammalian and yeast cells or in unglycosylated form from prokaryotes. Recombinantly produced human IL-3 is purified in a series of steps, initially employing hydrophobic interaction, followed by ion exchange chromatography and gel filtration.

Abstract: The present invention provides a method for purification of a surface exposed, immunogenic outer membrane protein of Haemophilus influenzas which is conserved amongst strains. The protein, designated P6, is relatively free of detergent, contaminating RNA and undesirable cellular components.In accordance with the present invention, there is provided a method for purifying an immunogenic outer membrane protein of H. influenzas consisting essentially of:a) suspending H.

Abstract: There is provided in accordance with the practice of this invention a process for separating Factor VIII complex from an impure protein fraction containing Factor VIII complex. An aqueous solution of the impure protein fraction containing Factor VIII complex is applied to a heparin-coupled chromatographic medium, to bind the Factor VIII complex to the medium. The Factor VIII is then recovered from the heparin-coupled chromatographic medium by elution with an aqueous solution comprising CaCl.sub.2 and histidine. The recovered Factor VIII is further purified by precipitation with a solution comprising glycine and NaCl, and washing the resultant precipitate with a solution comprising histidine, glycine, and NaCl to provide a Factor VIII complex solution with a specific activity of about 70 to about 150 units/mg.

Abstract: A method of isolating cystine rich acid-stable, biologically active proteins comprising the steps of co-precipitation by acidification of said proteins together with at least one acid-sensitive protein; isolation of said proteins from the co-precipitate by its resuspension in aqueous solution and the subsequent recovery of the proteins from the supernatant.

Abstract: The invention proposes a process for purifying a highly glycosylated protein from a crude preparation which comprises the action (i) of adding to said preparation a divalent metal ion in a sufficient amount in order to form a mixture which precipitates and (ii) after precipitation, of harvesting said protein from the mixture supernatant.

Abstract: A method of preparing an IgA rich preparation comprising exposing a plasma fraction to an amino acid, organic salt or inorganic salt with optional chromatographic treatment yielding a product suitable for use in medical conditions treatable with IgA.

Abstract: This invention describes processes for producing mature human members of the NGF/BDNF family of neurotrophic proteins that are fully biologically active. In addition, the gene encoding human BDNF and processes for obtaining the same are disclosed.A previously-unreported member of the NGF/BDNF family of neurotrophic proteins, NGF-3, has been identified and a portion of the gene encoding for the NGF-3 has been described. Processes for identifying additional previously unreported members of the NGF/BDNF family are also described.

Abstract: Immunogenic components are recovered from pathogenic bacteria grown on semi-defined, serum-free media in purified forms that are useful as veterinary acellular vaccines and include both polysaccharides as well as proteins. Recovery is accomplished by homogenizing the cells; precipitating unwanted cellular compounds such as nucleic acids using a quaternary ammonium salt to form insoluble ionic complexes; treating the resulting supernatant with a hypersaline solution to dissociate any residual ionic complexes; and concentrating and dialyzing the supernatant to remove the ammonium and chloride salts and various unwanted components derived from the culture medium.

Abstract: A method is provided for recovering astaxanthin, astaxanthin carotenoids, astaxanthin esters, chitin and proteins from crustacean tissues containing such. The method comprises an initial extraction of crustacean tissue with boiling lye to form an alkaline extract and an extracted residue. The alkaline extract, upon cooling, forms separate layers from which can be recovered component protein, astaxanthin, astaxanthin carotenoids and astaxanthin esters. The lye extracted residue of chitin-containing such crustacean tissue is readily processed to provide chitin.

Abstract: Methods for purifying factor XIII from a biological fluid are provided. The methods comprise precipitation of factor XIII by adjusting the pH of the biological fluid to 5.5 to 6.5 and recovering the precipitated factor XIII.

Abstract: A process for the preparation and purification of recombinant Interferon-.alpha. is disclosed. The invention is directed to a process comprising the following steps: cultivating E. coli containing the interferon gene for a growth period during which not more than 5% methionine-interferon is formed; extracting and concentrating the expressed interferon; subjecting the preliminarily purified material to Tandem Chromatography, wherein the Tandem Chromatography comprises separation on a cellulose column followed by an anti-alpha-interferon monoclonal antibody affinity column; subjecting the thus purified material to isoelectric precipitation of impurities at about pH 4.0 to about pH 4.8; and purifying the interferon by chromatography on a high performance cation exchange column using a volatile buffer, wherein the purified interferon is non-immunogenic when administered parenterally to a human.

Abstract: A process is disclosed in which high purity protamine-DNA complexes are prepared by collecting nucleoprotamines specific developmental stages of a life form, specifically, amphibian, egg by low temperature processing. The process also includes the steps of sequential homogenization in a high concentration aqueous salt solution at a buffered low pH, followed by ultracentrifugation to remove insoluble matter. Either a crude mixture or pure isolate of the complexes may be produced. Pure isolates require aqueous chloroform extraction to isolate protein and to remove lipids. Lyophilization then removes chloroform and excess water. The isolate is then fractionated by single pass alumina chromatography. Dialysis against pure water removes salts. Repeated lyophilization removes excess water and concentrates single protamines and protamine-like proteins. The mixture may then be reconstituted with 5% weight/volume heterologous or homologous DNA, in order to shield from charge toxicity.

Abstract: A multi-step process for purifying an immune serum globulin fraction from a crude plasma protein fraction involves precipitating non-serum globulin proteins from an aqueous suspension of the crude plasma protein fraction using a protein precipitant, adding a virus-inactivating agent to the clarified immune serum globulin-containing liquid, absorbing the immune serum globulins onto a cation exchange resin and washing non-serum globulin contaminants from the resin, subjecting the eluate to ultrafiltration to concentrate the immune serum globulins and separate them from low molecular weight species, contacting the concentrate with an anion exchange resin to absorb non-serum globulin contaminants, passing the imune-serum globulins through the anion exchange resin under conditions that leave non-serum globulin contaminants bound to the resin, and subjecting the filtrate to a molecular washing step to produce a purified immune serum globulin fraction.

Abstract: A process for the purification of proteins from solutions containing contaminants of similar net charge and molecular weight is provided, comprising contacting a solution containing the desired protein with an immobilized metal affinity chromatography resin in a buffer containing a low concentration of a weak ligand for the chelant of the resin. The adsorbed protein is then eluted using a buffer having a high concentration of the same weak ligand, e.g., Tris. Particularly preferred features employ agarose-iminodiacetic acid resins having copper cations and are especially useful in obtaining preparations of homogeneous, stable rsT4 proteins.

Abstract: The invention relaates to the production of collagen fibers by comminuting collagen containing tissues, drying the comminuted product and milling the dried material while maintaining the temperature sufficiently low to prevent substantial conversion of collagen to gelatin. The collagen fiber product is particularly useful for restructuring poorly textured meats, mechanically recovered meat products, offal, fish, fish products and other protein products to improve textural properties, water retention, fat retention, eating quality, juicines, succulence, shape, size retention and protein content.

Abstract: A process for recovering highly pure, recombinant IL-2 from transformed microorganisms in which the cells are disrupted; impure recombinant IL-2 is isolated in the form of refractile bodies from the disruptate; the impure IL-2 is dissolved and denatured with at least 6M guanidine hydrochloride containing a reducing agent; the reduced IL-2 is precipitated and resolubilized; the reduced solubilized IL-2 therein is oxidized by a controlled oxidation; the oxidized IL-2 is refolded by reducing the concentration of guanidine hydrochloride in the solution; and the oxidized, refolded IL-2 is further purified by ion exchange chromatography or hydrophobic interaction chromatography and ion exchange chromatography.

Abstract: An essentially pure and stablized antibody preparation comprising IgM antibodies having a purity greater than about 98% by weight and a nucleic acid content of less than about 200 pg per mg IgM. In one embodiment IgM antibodies from a monoclonal source are subjected to ion exchange and size exclusion chromatography at an alkaline pH to yield a purified IgM having a nucleic acid content of less than 10 pg/mg IgM, preferably less than about 4 pg/mg IgM. A highly purified and stabilized preparation of anti Pseudomonas aeruginosa antibodies is disclosed. The removal of nucleic acids is assured by subjecting the antibody source to at least one and preferably two low pH precipitation steps. In a very preferred embodiment, ion exchange and/or size exclusion chromatography is used to remove any residual nucleic acids.

Abstract: A substantially heme-free blood protein product is produced by adding an acid to a blood cell suspension to a low pH to disrupt erythrocytes present in the suspension thereby releasing hemoglobin, and to cleave off the heme moiety from the major proportion of the hemoglobin. The released heme forms a precipitate which is removed by centrifugation. Heme moieties remaining in the blood protein solution are degraded by treatment with an oxidizing agent, e.g. hydrogen peroxide. The product has a content of sulfur-containing amino acids which corresponds to the content thereof in natural hemoglobin, as well as foaming and emulsifying properties corresponding to naural hemoglobin. The product has a high lysine content and may be used to enrich the protein content of edible products, e.g. cereals, or to replace part of the meat in meat products. An edible product containing up to about 25% by weight of blood plasma mixed with the product is also disclosed.

Abstract: A process for producing the protein part of Ca.sup.+2 -binding photoprotein aequorin (apoaequorin) according to a recombinant DNA technique, and a process for purifying the resulting apoaequorin are provided, which production process comprises cultivating a strain having an expression vector piP-HE outside the bacterial bodies transformed into Escherichia coli, followed by separating the resulting culture solution into the bacterial bodies and a culture filtrate, and recovering the culture filtrate, and which purification process comprises adding an acid to the culture filtrate so as to give a pH of 4.7 or less, followed by recovering the resulting white precipitates, dissolving the white precipitates in a buffer solution, reducing the solution, subjecting the resulting apoaequorin fraction to adsorption treatment according to anion exchange chromatography and subjecting the resulting apoaequorin to gel filtration.

Abstract: A method of extracting granulocyte/macrophage colony stimulating factor (GM-CSF) from GM-SCF-expressing bacterial cells comprising treating a suspension of GM-CSF-containing bacterial cells with an acid and an enhancing agent, or with an acid that is itself an enhancing agent, removing substantially all of the suspension liquid from the cells, preparing a second suspension of the acidified cells, neutralizing said second suspension, and separating the GM-CSF-containing liquid from the suspended cells.

Abstract: This invention provides a crystalline form of recombinant human granulocyte-macrophage colony-stimulating factor (r-h-GM-CSF) and methods for making such crystals.

Abstract: A method for recovering a recombinant protein from a protein solution containing high molecular weight containing proteins by directly adding amine or quaternary ammonium compounds to the solution in amounts sufficient to selectively precipitate the high molecular weight protein contaminants.The high molecular weight precipitates are removed and the solution is further processed to remove low molecular weight contaminating proteins and other non-protein contaminants. The recombinant protein is subsequently recovered and further processed to produce a protein composition suitable for its intended use.

Abstract: A method of preparing a sterile and stable plasma-protein solution containing fibrinogen and Factor XIII from human blood plasma which has been stabilized with citrate, comprising treating the plasma with .beta.-propiolactone and irradiating it with ultraviolet light, removing the Factors II, VII, IX and X by adsorption onto anion exchangers that adsorb proteins, precipitating the companion proteins out by adding ethanol until the solution has a final concentration of about 9% by volume at -3.degree. C., centrifuging the precipitate off, dissolving the precipitate in a citrate buffer at a pH of about 6.35 and a temperature of about 37.degree. C., adjusting the protein level of the solution to about 13.3 g/l with sodium citrate solution, adding ethanol, a glycine citrate buffer, and a solution of sodium citrate to precipitate out the companion proteins, adding ethanol to the remaining solution until the solution has a final concentration of about 9% by volume at -3.degree. C.

Abstract: An immunoglobulin-rich fraction containing IgG, IgA and IgM is prepared in high concentration (200 mg/ml) by contacting diluted serum with CuSO.sub.4, which is a source of strong positive ions, to produce a precipitate rich in immunoglobulins, whereafter the precipitate is washed with an EDTA chelating agent solution and then a solution of L-lysine.HCl and NaHCO.sub.3 is added to form a colloidal solution of the washed immunoglobulin precipitate.

Abstract: A blood substitute and plasma expander comprising a cross-linked, substantially endotoxin-free homoglobin solution and process for preparing same. The process comprises fractionating whole blood, separating out a stromal-free, sterile hemoglobin solution, chromatographically separating endotoxins from said hemoglobin solution and crosslinking the resulting endotoxin-free hemoglobin solution.

Abstract: This invention deals with a simple and efficient method for producing vitronectin from biological materials which include vitronectin. More specifically, this invention deals with a method for producing vitronectin by binding vitronectin in biological materials to immobilized glycosaminoglycans in the presence of protein denaturing agent, especially urea.

Abstract: A preparation of an isolated immunoreactive CA 125 Antigen, and a method of isolating it is disclosed. CA 125 Antigen is a glycoprotein having a molecular weight of about 200kD, and a carbohydrate-content of about 24%. The CA 125 Antigen is isolated from a cell culture medium by acid precipitation, and is subsequently purified by size exclusion chromatography and immunoaffinity chromatography.

Abstract: A method of removing a constituent of a biological fluid including a blood component, said method including flowing the biological fluid past one side of a first semipermeable membrane, flowing solution containing a first precipitation agent past a second side of the membrane so as to cause transfer of the precipitation agent through the membrane to the biological fluid so as to improve precipitation characteristics of the fluid; and precipitating the constituent from the biological fluid. Also disclosed are maintaining a lower pressure in a biological fluid in a dialyzer than in dialysate at all portions of a membrane in the dialyzer and adding a continuously flowingy stream of concentrated precipitation agent to a continuously flowing stream of a biological fluid.

Abstract: A process for purifying human G-CSF from an aqueous solution including the step of adding NaCl to the solution to selectively precipitate the human G-CSF.

Abstract: A process for isolating and purifying GM-CSF produced from recombinant sources. The process provides for the isolation and purifying of recombinant GM-CSF produced in E. coli.

Abstract: A method for recovering a recombinant protein from a protein solution containing high molecular weight contaminating proteins by directly adding a flocculant to the solution in amounts sufficient to selectively precipitate the high molecular weight protein contaminants is disclosed.The high molecular weight precipitates are removed and the solution is further processed to remove low molecular weight contaminating proteins and other non-protein contaminants. The recombinant protein is subsequently recovered and further processed to produce a protein composition suitable for its intended use.

Abstract: This invention relates to a process for continuously fractionating plant, animal or human proteins by selective precipitation of the proteins resulting from placing a solution of proteins in contact with a precipitating agent constituted by a fatty acid of 6 to 14 carbon atoms, such as caprylic acid, is characterized in that respective deliveries of fatty acid and of the protein solution are continuously placed in contact in a mixing chamber of small volume with respect to the deliveries, creating a strong stirring in this mixing chamber; the individual deliveries of fatty acid and of protein solution are adjusted to controlled pH and temperature so as to maintain their ratio equal to a predetermined value; the mixture is then allowed to evolve during a phase of maturation so as to form a suspension; this suspension is separated into a liquid part from which are extracted the proteins having remained soluble, and a solid part containing proteins of different nature; and the parameters intervening in the proce

Abstract: A process for the solubilization of a protein obtained via heterologous expression comprises the step of bringing the insoluble protein produced in transformed host cells into contact with an aqueous acid solution in the presence of a protein denaturant. The process also preferably comprises the further steps of elevating the pH value of the solution, by addition of alkali substance, and thereafter lowering the pH to obtain a biologically-active protein.

Abstract: Recombinant hepatitis B antigen bound to yeast membranes in yeast expression systems is rapidly purified by subjecting the membrane bound protein to agents that release undesired proteins, followed by agents that release the recombinant hepatitis B antigen.

Abstract: A ciliary neurotrophic factor (CNTF), particularly sciatic nerve CNTF (SN-CNFT) is claimed. The SN-CNTF described herein is a single protein species and has a specific activity that increased to greater than 25,000-fold from crude extract. The purification is carried out by lowering the pH of the crude nerve extract preparation to form a precipitate which is removed and discarded; raising the pH to about 6.3 followed by ammonium sulfate fractionation; chromatofocusing a solution containing a second precipitate obtained from the 30% to 60% ammonium sulfate containing solution; subjecting the fractions obtained by chromatofocusing to sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE); and performing reversed-phase high-performance liquid chromatography (HPLC) on the SDS-PAGE eluate.Amino acid data for this SN-CNTF is also provided. In addition, methods for using this data for providing SN-CNTF probes and for screening cDNA and genomic libraries are also provided.

Abstract: Methods of purifying recombinant surface antigen of hepatitis B virus are disclosed. In one protocol, purification is achieved by selective extraction of the antigen from yeast membranes, followed by solubilization with urea and dithiothreitol.

Abstract: A process for the preparation of a single crystal of a biopolymer by growth from a solution, which comprises continuously changing one factor having an influence on the conditions for crystallization of a solution of a biopolymer, fractionating the solution, and independently crystallizing the resultant fractions. The process may be carried out by an apparatus comprising a means for feeding a crystallizing agent solution, a means for feeding a bioploymer solution, a means for producing a series of changes of predetermined crystallization conditions, said means continuously changing at least one factor having an influence on the conditions for crystallization of the biopolymer solution, and a means for fractionating the solution and independently crystallizing the resultant fractions.

Abstract: A method for recovering a recombinant protein from a protein solution containing high molecular weight contaminating proteins by directly adding Group IIA metal salts to the solution in amounts sufficient to selectively precipitate the high molecular weight protein contaminants is disclosed.The high molecular weight precipitates are removed and the solution is further processed to remove low molecular weight contaminating proteins and other non-protein contaminants. The recombinant protein is subsequently recovered and further processed to produce a protein composition suitable for its intended use.

Abstract: Disclosed is a method of recovery of antihemophilic factor (AHF) from human plasma by precipitation with citric acid, sodium citrate, or potassium citrate.

Abstract: A method of removing a constituent of a biological fluid including a blood component, said method including flowing the biological fluid past one side of a first semipermeable membrane; flowing solution containing a first precipitation agent past a second side of the membrane so as to cause transfer of the precipitation agent through the membrane to the biological fluid so as to improve precipitation characteristics of the fluid; and precipitating the constituent from the biological fluid. Also disclosed are maintaining a lower pressure in a biological fluid in a dialyzer than in dialysate at all portions of a membrane in the dialyzer and adding a continuously flowing stream of concentrated precipitation agent to a continuously flowing stream of a biological fluid.

Abstract: A method is provided for extracting human interleukin-4 (IL-4) from IL-4 expressing bacterial cells. The method comprises treating the bacterial cells with a deactivating agent, disrupting the cell membrane and recovering the IL-4.

Abstract: A method of recovering lactose from milk whey or cheese whey with a high yield, wherein the whey is concentrated, part of the lactose is crystallized, and the crystals are separated and dried; the mother liquor is purified by heating it to about 60.degree. to 70.degree. C. at a pH of about 5.8 to 7.0 and by removing the resultant precipitate by centrifugation; the purified mother liquor is fractionated chromatographically, using sulphonated polystyrene resin which is in sodium ion form and cross-linked by a 3-6% per weight divinylbenzene and which is even-grained, the flow rate is about 0.4 to 1.5 m.sup.3 /h, the temperature about 50.degree. to 75.degree. C., and the pH about 5.5 to 7; whereafter elution is carried out with water, and the following fractions are recovered: a protein fraction which may also contain salts, an intermediate fraction which is recirculated to the fractionating step, and a lactose fraction which is passed to the crystallization step.

Abstract: A process for recovering dimeric, biologically active CSF-1 from bacterially expressed recombinant CSF1 genes is described. The process comprises recovery of the solubilized monomeric form, followed by dimerization under refolding conditions and further purification of the dimer. Heterodimers may also be produced by this process.

Abstract: Biologically active lymphotoxin polypeptide species, and derivatives, fragments, aggregates and pharmaceutically acceptable salts are provided. The lymphotoxins are substantially homogenous, and are formulated into pharmaceutical compositions. The lymphotoxins are purified to a specific activity of at least 10.sup.6 units/mg protein by using hydrophobic substances and/or immobilized lentil lectin, or with the use of other chromatographic processes such as ion exchange chromatography, HPLC or gel filtration.

Abstract: Process of crystal growth by diffusion, particularly intended to be carried out on board a space vessel and according to which a substance (5) to be crystallized, contained in a crucible (4), is brought into presence with a precipitating agent (9) through a diffusion wall (6) obturating said crucible. According to the invention:in a first recipient (1) is arranged said crucible (4) containing the substance to be crystallized and obturated by said wall;said first recipient (1) is obturated by a stopper (3) of porous material;in a second recipient (12) is arranged a porous body (19);said first and second recipients are brought together so that said porous body and said stopper of porous material are in contact with each other; andsaid precipitating agent is injected into said porous body (19) of said second recipient (12) through (15).

Abstract: A method of extracting granulocyte macrophage colony stimulating factor (GM-CSF) from GM-CSF-expressing bacterial cells comprising treating a suspension of GM-CSF-containing bacterial cells with an acid and an enhancing agent, removing substantially all of the suspension liquid from the cells, preparing a second suspension of the acidified cells, neutralizing said second suspension, and separating the GM-CSF containing liquid from the suspended cells.

Abstract: Interleukin 2 (IL 2; T-cell growth factor), produced with and without costimulation by Burkitt's lymphoma line Daudi, is highly purified approximately over 37,000-fold to apparent homogeneity from lymphocyte-conditioned medium derived from normal humaThis invention was made with support in part under Grants CA 08748, CA 22507, CA 25608, CA 20194, CA 21525, CA 31525, P01-CA-20194, AI 18 321-01, CA 23766 and CA 33050 awarded by the National Cancer Institute, National Institute of Health, DHEW. The government has certain rights in this invention.

Abstract: Basic Fibroblast Growth Factor (FGF) is substantially purified by the employment of affinity chromatography using heparin-linked support material. Described is a simplified three step procedure for extracting basic FGF from either mammalian brain or mammalian pituitary tissue. Salt precipitation, e.g., with ammonium sulfate is used to provide a partially purified precipitate that is then subjected to ion-exchange chromatography, e.g., using a Carboxymethyl-Sephadex column. Substantially pure basic FGF fractions are then obtained by fractionating the further partially purified fractions using affinity chromatography on a heparin-linked support e.g., Heparin-Sepharose.

Abstract: A liquid phase containing preS2+S antigen is separated into two phases after which the desired antigen is concentrated, washed and adsorbed and desorbed from a fumed silica to produce a product that is purified and concentrated in antigen:protein ratio and that is suitable for final purification.

Abstract: A method for the solubilization and recovery of insoluble parasite protective antigenic factors associated with parasite material comprising solubilizing the antigenic factors with a non-ionic detergent and separating the solubilized material from undispersed residual material. The purified protective antigenic factors are useful as vaccines, particularly against malaria, and as diagnostic agents.