Abstract: Improved methods for direct purification of product immunoglobulins or their derivatives from large volumes of mammalian cell culture medium include directly subjecting the cell culture medium to cation exchange treatment, so as to adsorb the product but not the contaminants. The eluted product is then recycled, or is applied to anion exchange, for further purification, and optionally subjected to additional steps. The product may be obtained in a form suitable for clinical applications, if desired.

Abstract: A method of and apparatus for separating proteins adsorbed by ion-exchange gels, especially anion-exchanging gels includes at least one chromatographic column in which the protein-carrying gel is charged and is subjected to a gradient-elution with a buffer solution as eluant whose property is changed with time by gradually changing the ionic strength and maintaining a substantially constant pH value or by gradually changing the pH value and maintaining substantially a constant ionic strength. The obtained eluate is then fractionated into its various components.

Abstract: An agent for the therapy of hemophilia A which is resistant to treatment with factor VIII is described, and is obtainable by maintaining a mixture of factor VIII, antithrombin III, a phospholipid and calcium ions in an aqueous solution at a temperature of from 1.degree. to 45.degree. C. for at least one minute, adding factor IX, and maintaining the solution at a temperature offrom 1.degree. to 45.degree. C. until addition of a sample of this solution to an inhibitor plasma results in a partial thromboplastin time (PTT) of 15 to 30 seconds, where appropriate adding a polyol and, where appropriate, an amino acid, and, where appropriate, drying the solution.

Abstract: Immunoglobulin M is purified by absorption upon an insoluble matrix material having chemically bound thereon the protein C1q. The matrix is washed and purified immunoglobulin M is then released from the matrix.

Abstract: A simple, rapid and reliable assay for measuring functional enzyme inhibitor levels in body fluids and tissues, especially functional .alpha..sub.1 -proteinase inhibitor levels in human plasma or serum. .alpha..sub.2 -Macroglobulin is first inactivated, then porcine pancreatic elastase incubated with the samples to form a complex between the elastase and the functional .alpha..sub.1 -PI. Deficient individuals are detected by the presence of a color change following addition of substrate. If desirable, residual enzyme activity can then be calculated and the .alpha..sub.1 -PI levels present in the original sample determined. The method provides a means for early screening of individuals with a genetic deficiency in circulating levels of .alpha..sub.1 -PI, thereby facilitating treatment and prevention of familial emphysema.

Abstract: A method for the separation of blood into the components thereof, characterized by subjecting the blood to strong centrifugation thereby separating said blood into an upper layer (A) of platelet-deficient blood plasma an intermediate layer (B) of a mixture of platelets and white blood corpuscles, and a lower layer (C) of a red blood corpuscle concentrate, adjusting the hematocrit value of said red blood corpuscle concentrate of (C) to not more than 80% by diluting said red blood corpuscle concentrate with part of said platelet-deficient blood plasma of (A), and subjecting said mixture of (B) to weak centrifugation thereby separating said mixture into a lower layer of white blood corpuscles and an upper layer of a platelet concentrate, and if desired, subjecting the remaining platelet-deficient blood plasma to the treatments of freezing, thawing, and centrifugation thereby separating said platelet-deficient blood plasma into cryoprecipitate and cryoprecipitate-deficient blood plasma, and apparatus therefor.

Abstract: Anti-thrombobenic, anti-microbial compositions containing heparin reacted with quaternary ammonium components and bound with water-insoluble polymers are disclosed. Such compositions may also contain additional quaternary ammonium compounds not reacted with heparin and may also contain quaternary ammonium compound(s) reacted with antibiotics.

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 method of recovering active, highly purified and concentrated vitamin K-dependent proteins from plasma, concentrate or mixtures of proteins produced by recombinant DNA technology using an immuno adsorbent comprising a monoclonal antibody.

Abstract: A process for the production of a preparation of blood coagulation factor VIII which makes it possible to obtain a pasteurized product which is virtually free of immunoglobulins, isoagglutinins, fibronectin and coagulable fibrinogen is described.A product of this type can be used for the treatment of blood coagulation disturbances.

Abstract: Hepatitis B PreS2+S antigen is isolated from plasma by adsorption on an affinity chromatography column, elution with a chaotropic agent and treatment with concentrated urea at an elevated temperature. The process retains all or substantially all of the preS2+S antigen.

Abstract: A method and therapeutic composition for the treatment of bleeding disorders, for example those characterized by a tendency toward hemorrhage or a hypercoagulative state, by the administration of tissue factor protein or antagonists thereof.

Abstract: An improved assay method for detecting the presence of an antibody capable of binding with an antigen of a virus is provided. The improvement comprises using a non-human immune antibody which is reactive with an anti-human antibody as a positive control in the assay. Non-human immune IgM and a method of producing the IgM is also provided.

Abstract: Substantially pure cromolyn binding protein is prepared by means of affinity chromatography of cromolyn derivatives bound to insoluble matrices. Aminocromolyn is prepared by a six-step synthesis and amine derivatives thereof are prepared by conventional means. Obtaining a compound having an amine group instead of the OH group at the 2-carbon of the propane link of cromolyn permits many kinds of reactions without interfering with the portion of the cromolyn molecule with causes its pharmacological activity. The cromolyn derivatives can be conjugated to proteins such as BSA by means of glutaraldehyde cross-linking and such conjugates can be covalently bound to agarose beads. Cromolyn binding protein can be isolated by passing lysates of RBL-2H3 cells through chromatographic columns packed with such beads. The cromolyn binding protein can be further purified by means of lectin-agarose columns.

Abstract: A novel class of compounds, methods for the preparation thereof and the use thereof in chromatographic methods for binding various biologically active materials non-covalently are disclosed. The class of compounds comprises the reaction product of a polymeric gel with a pyridine base, such as 4-dimethylaminopyridine (DMAP), and a halogen-substituted pyridine, such as 3,5-dichloro-2,4,6-trifluoropyridine (DCTFP), which reaction product may in turn be optionally reacted with hydroxyl ions or specified low-molecular-weight compounds. These compounds are capable of selectively and efficiently binding proteins and other organic materials of interest non-covalently to a degree comparable or superior to the heretofore preferred natural affinity ligands, such as Protein A gels. The novel compounds find particular utility in purification and recovery of proteins such as serum albumin and immunoglobulins of various classes from crude sources, such as diluted serum samples from various species.

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 of sterilizing plasma or plasma fractions, including fractions that contain the blood-coagulating Factor VIII by treatment with .beta.-propiolactone or ultraviolet radiation. Treatment with tri-n-butyl phosphate and sodium cholate or Tween 80 is carried out either prior to or simultaneously with the .beta.-propiolactone treatment or ultraviolet radiation.

Abstract: A set for the isolation of cryoprecipitate includes a hollow vessel with a closed first end, a second end and a longitudinal axis, and a nipple extending from and closing the second end, said nipple enclosing a volume of approximately 2 to 5 percent of the volume of the hollow vessel. In one embodiment, the vessel is formed of a semi-rigid material and the first end is closed by a cap containing a micro-porous filter for venting the vessel. In another or further embodiment, the nipple has a tapered tip portion which may be sliced off to permit extrusion of the isolated cryoprecipitate by squeezing the vessel. In a different further embodiment, the nipple has a twist-lockable connector, for attachment to a syringe or to an applicator tip. In yet another embodiment a piston mounts on the cap within the vessel for directly extruding separated cryoprecipitate from the nipple.

Abstract: A set for the isolation of cryoprecipitate includes a hollow vessel with first and second ends and a tapered body extending to a narrow sump closing the second end, said sump enclosing a volume of approximately 2 to 5 percent of the volume of the hollow vessel. A port enters the vessel in the tapered body above the sump. In one embodiment, the vessel is formed of a rigid or semi-rigid material and the first end is closed by a cap containing a micro-porous filter for venting the vessel. In another or further embodiment, the sump has a tapered tip portion which may be sliced off to permit extrusion of the isolated cryoprecipitate by squeezing the vessel. In a different embodiment, the vessel contains an inner filter column, or bag with a filter matrix, accessible via an access port. Cellular material placed in the column is frozen, thawed and centrifuged to separate out antigen-free platelet growth factor with the cryoprecipitate.

Abstract: A megakarytocyte stimulatory factor (MSF), purified to homogeneity, is an acidic protein (pI=5.1) with an Mr=15,000 which stimulates PF4-like protein synthesis in rat promegakaryoblast cells by as much as 7-fold, and exhibits half-maximal activity at a concentration of 0.8 pM. MSF exhibits no biologic activity corresponding to other known hemopoietic growth factors, and appears to be specific for the megakaryocyte lineage.In the given examples, MSF was purified to homogeneity (as judged by SDS-PAGE and isoelectric focusing in the presence of 9.2 M urea) from serum-free conditioned medium obtained from cultured human embryonic kidney (HEK) cells, and to near homogeneity from thrombocytopenic plasma.

Abstract: Sensitive proteins and other macromolecules, such as enzymes, antibodies, antigens, serum complement, fluorescent proteins, vaccine components, polysaccharides such as agarose etc, can be preserved by drying at ambient temperature and at atmospheric pressure in the presence of trehalose. A porous matrix impregnated with trehalose is provided as a receiver for a blood or other liquid sample to be dried.

Abstract: The administration of Factor I and/or Factor H to a mammal has been found to alleviate or prevent such autoimmune diseases as systemic lupus erythematosus, rheumatoid arthritis and glomerulonephritis.

Abstract: Novel, biologically active fragments of human antihemophilic factor, processes for their preparation, pharmaceutical preparations containing them and the use of such fragments in the treatment of patients suffering from hemophilia.

Abstract: A method for treatment of acquired immuno-deficiency syndrome (AIDS) which comprises administering a therapeutically effective amount of liposomes containing a toxin which specifically inhibits the protein synthesis in cells to a patient with AIDS.

Abstract: This invention relates to cosmetic, health- and body-preserving compositions of high biological value, optimizing the biological processes occurring in the skin cells and providing the most preferable function of the enzyme system of the cells connected with the age of the organism.The compositions of the invention contain in addition to the commonly used carrier and additive and/or filling materials and active ingredients, mineral waters of a native condition, medicinal waters and/or the mixtures thereof and/or the mixtures thereof with fermented or non-fermented plant juices and/or optionally inorganic materials playing the role of trace elements in the living organism as well as proteins.

Abstract: The selective appetite-regulating substances satietin and satietin-D had up to the present been prepared from human or animal blood serum or plasma by ultrafiltration, gel chromatography, treatment with trichloroacetic acid, affinity chromatography and optionally treatment with a proteolytic enzyme.According to the invention, satietin and satietin-D are obtained in a purified form by using immunoabsorption. Being glycoproteins, both satietin and satietin-D form antibodies (antisatietin and antisatietin-D, respectively) in the living organism and can be isolated from the blood and bound to a gel column. For the preparation of satietin and satietin-D, respectively, the human or animal blood plasma or serum is subjected to ultrafiltration and the ultrafiltrate, containing in addition to satietin and satietin-D other constituents with a molecular weight below 50000 daltons, is contacted with the gel column.

Abstract: The invention relates to Tetranectin, a new protein isolated from blood. Its structure comprises four polypeptide chains having the formula shown in the attached drawing.Tetranectin plays a role in the hemostatic system and, therefore, may be used as an agent for regulation of hemostasis.Further, the invention relates to a process for preparing Tetranectin in which Tetranectin is isolated e.g. from blood or blood fractions, cells or genetically engineered organisms.Finally the invention relates to antiserum or antibodies against Tetranectin, to immunological detection and assay methods wherein said antiserum or antibodies are used as immunological reagent, and to pharmaceutical compositions containing Tetranectin or antibodies against Tetranectin.

Abstract: A process for heat treating an aqueous solution containing chemically unmodified .gamma.-globulin, wherein said heat treating is carried out in the presence of sorbitol is disclosed. By the heat treatment, impurity viruses can be inactivated without causing impairment of activities of chemically unmodified .gamma.-globulin, increasing polymer contents or increasing an anticomplement titer.

Abstract: Parenteral solutions comprising(a) a sparingly soluble medicament active compound,(b) a solvent consisting of(i) 5-100 M/V % of an organic solvent or of a mixture of organic solvents and(ii) 0-95 W/V % of water,(c) A 0.5-30 W/V % strength aqueous solution of a human serum protein and customary auxiliaries and/or excipients, 1 to 40,000 parts by weight, preferably 25 to 30,000 parts by weight of solvent (b) and 1 to 1,000,000 parts by weight, preferably 50 to 40,000 parts by weight, of human serum protein solution (c) being present per part by weight of medicament active compound.The sparingly soluble medicament active compounds which can be used have a solubility in water of between 1 .mu.g and 10 g, preferably between 10 .mu.g and 1 g per liter of water. Examples of such medicaments are dihydropyridine compounds and pyrazolones, and muzolimine.

Abstract: A new improved glycoprotein having erythropoietin-type activity is disclosed. The substrate is characterized by amino acid sequence substantially identical to the amino acid sequence of native human erythropoietin wherein the methionine-54 is replaced with leucine. DNA encoding for the EPO-substance and expression vectors incorporating the same are disclosed. Therapeutic compositions and methods for treatment of anemic conditions are described.

Abstract: The present invention is a protein purification column comprising an organic substrate matrix having low reactivity to proteins, said matrix being capable of maintaining monoclonal antibodies attached thereto in an external configuration and preventing interaction with the protein to be bound to the antibody, and a monoclonal antibody attached to the substrate, the monoclonal antibody having a specific affinity for the protein to be isolated.The present invention also is a method for isolating and purifying specific protein from a solution, wherein1. Protein-specific monoclonal antibody is attached to the organic substrate matrix described above to form an antibody-substrate conjugate; and2. Protein to be isolated, in an appropriate buffer solution, is contacted with the antibody-substrate conjugate.An appropriate buffer may be applied to remove non-antibody bound contaminants, followed by an appropriate eluting agent to remove the protein from the monoclonal antibody.

Abstract: There is disclosed a method of producing a Factor VIII (AHF) containing fraction having a specific activity of at least 2.5 units of Factor VIII/mg protein as well as a portion of immunoglobulin G (IgG) of 10 mg/1000 units of Factor VIII at the most. Its risk of transmission of viral or bacterial infections is to be avoided or largely reduced. The method consists in that undesired proteins are at first precipitated from a Factor VIII (AHF) containing plasma fraction in the presence of SPS. The purified Factor VIII containing solution is treated with suitable salts or salt mixtures in order to obtain a Factor VIII containing precipitate. This precipitate is dissolved, lyophilized and finally heat-treated.

Abstract: Immunoglobins are purified by adsorption upon an immobilized protein A adsorbent using a buffer having a pH of 7.5 to 10 and containing a combination of monovalent cations and polybasic anions in a concentration of about 0.6M to 1.75M.

Abstract: To diagnose diseases in patients, a protein complex, RhC, is prepared from horse serum by precipitating a white powder from the serum at a pH of 5.5 and processing to remove lipids at a pH of 8.2 using Tris-HCl as the buffer. It includes two components associated together to provide a molecular weight of 280,000 and having characteristics of a rheumatoid factor and a Clq-like subcomponent of the complement. The protein complex is incubated with human serum or plasma and then precipitated by dialysis against a high pH buffer (0.05 M Tris-HCl pH 8.2). When precipitated, it co-precipitates the immune complexes from the human blood serum without substantial monomeric immunoglobulin to quantitatively isolate immune complexes from serum. Immunological assays then determine how much immune complex and what kind were in the serum.

Abstract: High purity albumin can be recovered in high yield from human plasma protein solution containing albumin by heat-treatment of the solution to denature and precipitate non-albumin proteins in a combination of specific conditions such as protein concentration of the solution, pH, heating time and temperature, the concentration of precipitant ammonium sulfate.

Abstract: Novel, biologically active fragments of human antihemophilic factor, processes for their preparation, pharmaceutical preparations containing them and the use of such fragments in the treatment of patients suffering from hemophilia.

Abstract: It has previously been shown that the serum from patients suffering from a wide range of cancers contains a cancer marker protein having the ability to release RNA from cell nuclei. This cancer marker protein is purified by taking the protein fraction precipitating between 30% and 50% saturated aqueous ammonium sulfate solution, dialyzing a solution of the protein fraction against TMK buffer, chromatographing the dialyzed protein on a molecular sieve and isolating the fraction having a molecular weight of about 60,000; and removing albumin. Injection of the purified protein into rabbits, preparation of serum from blood of the rabbits and absorption of the sera with normal plasma fraction produces at antibody specific to the cancer marker protein and therefore useful in a radioimmunoassay or ELISA test for a wide variety of cancers.

Abstract: There is provided, in accordance with practice of this invention, a process for separating Factor IX and/or Factor X from an impure protein fraction containing protein in addition to Factors IX and X. A silica resin coupled with a ligand capable of binding Factor IX and/or Factor X is provided. An aqueous solution of the impure protein fraction is applied to the ligand-coupled silica resin to thereby bind the Factor IX and/or Factor X to the resin. The Factor IX and/or Factor X is then recovered from the resin by elution.

Abstract: The present invention is directed to a method of producing growth factors for mammalian cells. More particularly, the method of the present invention comprises isolating mammalian inducer T-lymphocytes, and stimulating growth factor production by contacting these inducer lymphocytes with an antigen or a mitogen. The resulting growth factors are then separated from the material produced by the stimulated lymphocytes.

Abstract: Normal plasma from donors who have not been vaccinated with a varicella-zoster vaccine can be screened for higher than normal titers of naturally occurring antibody to varicella-zoster virus. Those plasmas with high titers of such antibody can be pooled and fractionated to give hyperimmune globulin. The product may be treated to render it suitable for intravenous injection. Patients with varicella-zoster infection or at risk to such infection, may receive the present product to raise serum titers of varicella-zoster antibody.

Abstract: A method for maintaining intact, non-degraded von Willebrand factor by preventing the action of calcium activated protease(s) responsible for degradation of the factor. The action of the calcium activated protease(s) may be avoided by removing the blood platelet source of the protease(s), by filtering or centrifugal separation, or by inactivating the protease with a chelating agent removing the calcium, by a protease inhibitor, or an alkylating agent.

Abstract: A rapid and simple process for purifying human, bovine and porcine procoagulant protein Factor VIII on a large scale using sequential high performance size exclusion chromatography under, first, low salt concentration conditions and, second, under high salt concentration conditions from reconstituted commercial Factor VIII:C (complexed Factor VIII) concentrate. The chromatographic separation is carried out on a high performance size exclusion chromatographic column packed with porous beads having a particle size of from about 13 to about 35 microns, pore diameters of from about 500 to about 2000 Angstroms and a pore volume of from about 1.0 to about 1.8 ml per gram. The first chromatographic separation is carried in a buffered aqueous solution using the buffered aqueous solution as an eluant. The low molecular weight constituents (impurities) are separated from Factor VIII and the high molecular weight constituents (impurities).

Abstract: A porous mineral support such as a porous mineral oxide coated with an aminated polysaccharide polymer has cationic characteristics and is capable of reversibly fixing thereto biological macromolecules. This material is employed in the separation and purification of said biologic maromolecules.

Abstract: What is disclosed is a method for isolating the cold insoluble globulin (CIG) from blood plasma concentrates containing factor VIII.

Abstract: Normal plasma from donors who have not been vaccinated with a varicella-zoster vaccine can be screened for higher than normal titers of naturally occurring antibody to varicella-zoster virus. Those plasmas with high titers of such antibody can be pooled and fractionated to give hyperimmune globulin. The product may be treated to render it suitable for intravenous injection. Patients with varicella-zoster infection or at risk to such infection, may receive the present product to raise serum titers of varicella-zoster antibody.

Abstract: There is disclosed an improved method for separating one of alpha-1-proteinase inhibitor (also known as alpha-1 antitrypsin) and antithrombin-III from an aqueous solution of plasma proteins containing the same, such as from Cohn Fraction IV-1, Cohn Fraction IV, reworks of Cohn Fraction IV and IV-1, Cohn Effluent II & III and Cohn Effluent I. The method includes the steps of first holding an aqueous solution of plasma proteins containing one of alpha-1-proteinase inhibitor and antithrombin-III in a relatively large volume of buffer solution as solvent and at a pH adjusted to be relatively basic when compared to conditions heretofore known, and at a temperature in the range of from 2.degree.-50.degree. C. for a period of about 0.2-24 hours. Following the above treatment, alpha-1-proteinase inhibitor and antithrombin-III are obtained by applying conventional techniques to the resulting solution. Accordingly, the solution is then mixed with a polyalkylene glycol, e.g.

Abstract: Production of a high purity concentrate of the antihemophilic Factor VIII (AHF) by precipitation of an aqueous solution of cryoprecipitate from blood plasma in a first step with such an amount of polyethylene glycol (PEG), preferably about 4% by weight, as will precipitate a substantial amount of the present fibrinogen, subjecting the fibrinogen-free solution to a second precipitation step with preferably about 12% by weight of PEG in the presence of a salting-in agent, such as an amino acid, in particular lysine or arginine, or a carbohydrate, and then recovering the precipitate with a concentrated content of the present Factor VIII. The obtained Factor VIII concentrate with a very low content of immunoglobulins and other plasma proteins has a solubility in an aqueous injection medium of 45 to 500 units/ml and a high specific activity of up to 50 units/mg protein.

Abstract: A method for producing intravenously injectable IgG comprising a particulate separation step, an ion exchange separation step and an affinity separation step, and the substantially pure, intravenously injectable IgG produced by the method.

Abstract: Cryoprecipitate for the production of factor VIII is extracted from frozen blood plasma by continuously circulating the plasma in a crushed state between the peripheral and central regions of an externally, steadily heated thawing zone, with thawed mixture of liquor and cryoprecipitate being removed from the lowermost region of the zone by way of a path including a weir configuration. Preferably: the zone is formed by an axially upright cylindrical vessel; circulation is effected by at least one helical blade coaxially rotatable in the vessel, which blade extends over the vessel heights, is radially narrow compared to the vessel, is close to the vessel wall, and scoops material upwardly; the outlet from the vessel is tangentially opposed to the blade rotation; and heating is by way of a liquid jacket with generally axial flow therethrough.

Abstract: Blood and proteinaceous blood products employed for their physiological and/or immunological properties are free of viable enveloped viruses by treatment with low levels of ozone, levels at which substantially all of the physiological and/or immunological activity is retained.