Abstract: A method of preparing a recombinant influenza vaccine using DNA technology is provided. The resulting vaccine is a multivalent, preferably trivalent, influenza vaccine based on a mixture of recombinant hemagglutinin antigens cloned from influenza viruses having epidemic potential. The recombinant hemagglutinin antigens are full length, uncleaved (HAO), glycoproteins produced from baculovirus expression vectors in cultured insect cells and purified under non-denaturing conditions. The recombinant vaccine can be developed from primary sources of influenza, for example, nasal secretions from infected individuals, rather than from virus adapted to and cultured in chicken eggs. The process for cloning influenza hemagglutinin genes from influenza A and B viruses uses specially designed oligonucleotide probes and PCR. In the preferred embodiment, the cloned HA genes are then modified by deletion of the natural hydrophobic signal peptide sequences and replacing them with a new baculovirus signal peptide.

Abstract: Erythropoietin isoforms having a specific number of sialic acids per erythropoietin molecule are disclosed. Also disclosed are mixtures of such isoforms, pharmaceutical compositions containing such isoforms or mixtures thereof and methods of obtaining the erythropoietin isoforms.

Abstract: Methods for the purification of interleukin-12 (IL-12) is provided.

Abstract: The invention relates to a method for isolation of highly pure von Willebrand Factor in which recombinant von Willebrand Factor (rvWF) is chromatographically purified by anion exchange chromatography on an anion exchanger of the quaternary amino type in a buffer solution comprising buffer substances and optionally salt. The buffer solutions are preferably free of stabilizers, amino acids and other additives. According to this method, highly pure recombinant rvWF can be obtained, which is free from blood plasma proteins, especially free from Factor VIII, and is physiologically active. Further, the invention relates to a pharmaceutical preparation that contains rvWF, which comprises mulitimers with a high structural integrity.

Abstract: The present invention provides a method for eliminating the use of a displacer in displacement chromatography of proteins. Elimination of displacer is accomplished by producing an appropriate retained pH gradient using adsorbed buffering species. When the band velocity curves of the proteins under consideration intersect a vertical section of the pH profile and none of these protein have adsorption isotherm which cross each other at the pH of the intermediate plateau, and when the amount of protein in the feed slug to the column is such that bands of the appropriate concentration are formed in the displacement train, then a displacement pattern results in a chromatography column even though no displacer is present.

Abstract: Human growth hormone variants, DNA encoding the variants, vectors, host cells, pegylated forms of the variants, as well as methods of making the variants are disclosed.

Abstract: The invention provides methods for purification of human lactoferrin from milk, especially milk of nonhuman species, and for separation of human lactoferrin from undesired macromolecular species present in the milk, including separation from nonhuman lactoferrin species.

Abstract: Recombinantly produced serum albumin is purified in a series of steps, optionally by incubation with an anion-exchange adsorbent, followed by affinity chromatography employing a hydrophobic solid phase and using a water-soluble lipid anion as desorbens in the aqueous phase. This immobile phase comprises a carrier coupled to a 2-mercapto or 2-hydroxy alkanoic acid.

Abstract: Processes for isolating and purifying granulocyte colony stimulating factor (G-CSF) from a G-CSF producing microorganism are disclosed. The simplified processes include steps of lysing the microorganism and separating insoluble material containing G-CSF from soluble proteinaceous material; extracting the material with deoxycholate (optionally); solubilizing and oxidizing the G-CSF in the presence of a denaturant solubilizing agent and an oxidizing agent; removing the denaturant solubilizing agent from the G-CSF; subjecting the G-CSF to ion exchange chromatography; and recovering the purified G-CSF; yet excludes other cumbersome purification steps.

Abstract: A process for the isolation and characterization of a gene enzyme system for the inactivation of the herbicide phenmedipham, wherein the enzyme is a carbamate hydrolase of Arthrobacter oxidans, which is responsible for the cleavage of the carbamate bond between the benzene rings of phenmedipham. This process includes the isolation of the carbamate hydrolase, the identification of the amino acid sequence of two BrCN cleavage peptides of the carbamate hydrolase, the synthesis of oligonucleotides for specific determination of the carbamate hydrolase sequence by hybridization and identification of the coding region, cloning and specifying the nucleotide sequence of the carbamate hydrolase gene from Arthrobacter oxidans.

Abstract: The isolation and purification of type G botulinum neurotoxin and complexes thereof is disclosed. Compositions containing the type G neurotoxin and the preparation of a type G toxoid are also disclosed.

Abstract: The present invention relates to a new recombinant hybrid-DNA-molecule comprising a nucleotide sequence from S. aureus coding for a protein, or polypeptide, having fibronectin binding properties.

Abstract: The present invention generally relates to methods for purifying hemoglobin solutions and to hemoglobin solutions obtained by the methods. In one aspect, such methods include removing contaminants in crude hemoglobin-containing lysates with heat treatment. In a further aspect, the present invention provides methods for producing substantially purified hemoglobin solutions using immobilized metal affinity chromatography, optionally following by anion exchange chromatography.

Abstract: Porous zirconia particles of specific gravity of 2.5-3.5 g/cm.sup.3 and mean particle sizes of 30-400 .mu.m can be synthesized using oil emulsion methods from colloids and used for protein adsorption in stable expanded beds. Expanded beds of less than 1.0 settled bed height to diameter (approximately 10 ml bed volume) are stable at linear fluid velocities of at least about 100 cm/hour.

Abstract: The present invention relates to a purified casein phosphopeptide(CPP) having a novel amino acid sequence and a purified casein including same wherein the 25th Arg from N-terminal in a conventional CPP is replaced by Cys, rendering the CPPs to forming a dimer by disulfide bond. In the corresponding DNA sequence, cytosine is replaced by thymine to cause the amino acid replacement from Arginine(Arg) to Cysteine(Cys). The CPP or the casein containing same has an improved ability of solubilizing minerals and absorbing thereof in animals. The CPP or the beta-casein H containing same can be added to foodstuffs, beverages, medication, cosmetics, feed in an effective amount of enhancing a mineral absorption in animals. An oral composition comprising the beta-casein H or the inventive CPP and a pharmaceutically acceptable carrier can reduce or relieve a dentinal hypersensitivity.

Abstract: A process for purifying apolipoprotein A (ApoA) or apolipoprotein E (ApoE), or variants or mixtures thereof, comprises contacting a first aqueous solution comprising ApoA or ApoE and endotoxins with a matrix comprising an immobilized compound with an end group comprising two or three nitrogen atoms bonded to a carbon atom for attaching the endotoxins to the matrix, and subsequently treating the matrix comprising the immobilized compound with a second aqueous solution comprising a surfactant for releasing the ApoA or ApoE while the endotoxins remain attached to the matrix.

Abstract: A process for reducing degradation of recombinant coagulation factor VIII caused by metal-dependent proteases requiring Zn.sup.2+ for activity or containing Zn.sup.2+ as an integral part of their structure comprises adding an inhibitor of Zn.sup.2+ dependent proteases to a recombinant factor VIII solution. The recombinant factor VIII solution is obtained after harvesting a conditioned medium from a cell culture used for producing the recombinant coagulation factor VIII. The inhibitor is selected from complexing agents with a stronger affinity for the Zn.sup.2+ ion of the protease than for the ion or ions stabilizing the factor VIII molecule, and compounds structurally related to the natural substrate of the protease and containing an electronegative moiety.

Abstract: A process for the production of essentially homogeneous soluble, stable, endotoxin free human recombinant interleukin-1.alpha. of enhanced specific activity is described. The process involves breaking transformed microbial cells containing expressed human interleukin-1.alpha. and separating the soluble supernatant from the insoluble cell components and then passing the supernatant through gel chromatography and ion-exchange chromatography steps.

Abstract: Disclosed is a novel protein which has a molecular weight of 45,000.+-.5,000 and pI 5.7.+-.0.5 and exhibits cancer metastasis-inhibitory activity. The protein can be prepared by culturing human cells, animal cells and microorganisms capable of producing the protein in a nutrient culture medium while stimulating them with an inducer such as Bacille Calmette-Guerin and lipopolysaccharide.

Abstract: A method for preparing a purified and stable activated human Factor VIII composition is disclosed. A purified and stable activated human Factor VIII composition having a specific activity of at least 100,000 units per mg protein, or a potency of at least 15,000 units per ml is also disclosed.

Abstract: The invention concerns polypeptides with an apparent molecular weight of around 160 kD which are mediators or precursors for mediators of inflammation, derivatives thereof such as mutants and fragments, processes for their preparation, DNAs and hybrid vectors coding for the polypeptides and derivatives and host cells transformed with such hybrid vectors, polyclonal and monoclonal antibodies specific for the polypeptides or their derivatives and antibody derivatives as well as diagnostic and therapeutic methods for inflammatory conditions and Hodgkin lymphomas.

Abstract: A method for removing a prion from a solution comprising the prion and at least one additional biomolecule, comprising directing the solution through an anion-exchange chromatography column under conditions that cause a gradient elution, whereby the prion is separated from at least one of the biomolecules, thereby causing said biomolecule to be collected in an eluate fraction that is distinct from an eluate fraction that includes the prion. In one embodiment, the gradient is a pH gradient, for example, a step gradient. The prion can be a causal agent for a spongiform encephalopathy, such as Creutzfeldt-Jakob disease, Gerstmann-Straussler-Scheinken syndrome, scrapie, or bovine spongiform encephalopathy.

Abstract: Highly purified Pluripotent hematopoietic colony-stimulating factor (pluripotent CSF), a glycoprotein (MW 19,600) constitutively produced by human tumor cells has been highly purified from low serum-containing conditioned medium to apparant homogeneity. Pluripotent CSF supports the growth of human mixed colonies (CFU-GEMM), granulocyte-macrophage colonies (CFU-GM), early erythroid colonies (BFU-E) and induces differentiation of human leukemic cells. The specific activity of the purified pluripotent CSF in the CFU-GM assay is 1.5.times.10.sup.8 U/mg protein.

Abstract: The invention provides immunogenic oligosaccharide compositions and methods of making and using them. In particular, the compositions comprise oligosaccharides covalently coupled to carrier protein, wherein the resultant conjugate has been shown to contain specific immunogenic epitopes and elicits a protectively immunogenic response.

Abstract: A solid sorbent material comprising cellulose which has been modified by hydrolysis with a cellulase enzyme for a duration sufficient to increase the protein adsorption capacity of the solid sorbent material and methods for preparing the sorbent material. Methods for purifying a protein include passing a liquid medium containing the protein over the solid sorbent material are also included.

Abstract: The present invention relates to the method of detecting rupture of the amniotic membranes in pregnant mammals including humans using an immunoassay and reagents useful in such an assay. The method describes how to prepare a suitable protein antigen from amniotic fluid, gives criteria for the selection of this protein from the mixture of proteins present in amniotic fluid using the techniques of protein purification, gives criteria for assessing a sufficient degree of antigen purity for raising antibodies to the antigen and shows how the resultant antibodies can be used in immunoassays to detect the presence of amniotic fluid in the vagina and consequently to detect rupture of the amniotic membranes. The method also relates to the detection of amniotic fluid in other situations.

Abstract: A method of isolating a biomolecule from a medium containing biomolecules by ion exchange wherein the ion exchange material consists of a material having ion exchanging groups which can be transformed from a charged form to an uncharged form; the eluant comprises a charge neutralizing acid or base transforming the ion exchanging groups from the charged form to the uncharged form; and the charge neutralizing acid or base has a concentration in the eluant which is twice, preferably equal to or less than the concentration of the ion exchanging groups of the ion exchange material; the ion exchange material being in a packed, hydrated state. Preferably the method is for the isolation of phosphopeptides from a medium containing casein hydrolysates and the use of such isolated biomolecules is for the production of a food, a feed, a health care product, a cosmetic, or a pharmaceutical.

Abstract: Two or more analytes having the same action, or having different actions in spite of their similar structures, or two or more analytes having the same action and the same detectable chemical characteristics, in samples derived from living bodies, etc., can be measured rapidly and easily by forming one or more complexes with one or more affinity substances, separating the complexes by using high pressure liquid chromatography, followed by measurement of the amount of an affinity substance or one of the analytes.

Abstract: The invention concerns the removal of various allergologically irrelevant low-molecular weight components from the usual aqueous extracts of allergenically active proteins of plant pollens. Described are the desorption and subsequent elimination, from traditionally prepared allergenic pollen protein extracts, of low-molecular weight pigment and other compounds which are normally retained by strong electrostatic and/or hydrophobic forces. The preparation of such depigmented pollen proteins does not impair their allergenic potency or immunological specificity. The invention enables the production of fully active allergenic pollen proteins devoid of adhering low-molecular weight substances interfering with their safety, diagnostic accuracy and clinical efficacy. The purified pollen proteins represent improved starting materials for chemical derivatization, i.e. the preparation of attenuated vaccines for immunotherapy.

Abstract: The present invention relates to a purified non-neuronal activity-dependent neurotrophic factor (ADNF) protein that increase the survival of spinal cord neuron cells, cerebral cortical cells and hippocampal neuron cells which has a molecular weight of 16,000 to 18,000 Daltons as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and a basic pI of about 8.1.

Abstract: A method for purifying soluble laminin 5 from conditioned cell culture medium. A nonionic or anionic detergent is added to conditioned medium to a final concentration of between 0.1% and 1.0%. The conditioned medium is purified by cation exchange chromatography and anion exchange chromatography, yielding laminin 5 of at least about 70% purity.

Abstract: The invention provides a lipopolysaccharide (LPS) binding protein isolated from horseshoe crab. The LPS binding protein is isolated by (i) extracting the hemocyte membrane fraction of horseshoe crab with a polyethylene glycol ether type nonionic surface active agent in the presence of Ca ions, (ii) combining the extract with immobilized LPS under conditions that permit the LPS binding protein to bind the immobilized LPS to produce an LPS-LPS binding protein complex, and (iii) harvesting the LPS binding protein released from the complex in the presence of a chelating agent. The isolated LPS binding protein has a molecular weight of about 27,000 daltons as determined by SDS polyacrylamide gel electrophoresis and is operative to bind a lipopolysaccharide endotoxin. Accordingly, the isolated LPS binding protein can be used for detecting endotoxin and/or removing endotoxin from an injectable medicine.

Abstract: The invention relates to a process for the manufacture of very high-purity antihemophilic factor (FVIIIc) and von Willebrand factor. This process enables the manufacture of very high-purity antihemophilic factor (FIIIc) devoid of the bulk of the Willebrand factor comprises a step for purification by ion exchange chromatography with the aid of a chromatography column containing a gel, the purification step comprising a step for adsorption of the antihemophilic factor essentially devoid of the Willebrand factor on the gel in the column and a step for desorption of the purified antihemophilic factor, which is collected, thereby obtaining an antihemophilic factor devoid of the bulk of the Willebrand factor and having an activity as high as 250 IU/mg of proteins. This process also permits to recover von Willebrand factor in very high purity.

Abstract: Provided by the present invention are novel methods of protein recovery and purification methods and more specifically novel methods for the recovery and purification thioredoxin fusion proteins, especially of IL-11.

Abstract: A method is disclosed for the sequential separation of whey proteins using radial-flow chromatography. Different buffer systems adjusted to suitable pH and ionic strength are utilized in the separation process. The method separates at least five different proteins from whey. Infant feeding formulas, and other food formulations are also disclosed incorporating therein in different proportions various proteins separated from the whey.

Abstract: Methods of isolating components of interest from a milk sample are described. The methods include a step wherein the solubility of at least a portion of the total milk protein is stabilized in such a manner as to allow isolation of the component of interest without significant loss in yield. Kits for stabilizing the solubility of at least a portion of the total milk protein of the milk sample containing the component of interest also are described.

Abstract: A purified pneumococcal surface protein A (PspA) comprises a truncated form of the PspA protein which is immunoprotective and contains the protective epitopes of PspA. The PspA protein is soluble in physiologic solution and lacks at least the cell membrane anchor region of the whole protein. The protein is formed by insertion-duplication of mutagenesis of S. pneumoniae with pspA gene and expression of the truncated protein into the growth medium.

Abstract: The present invention provides isolated DNA molecules encoding human and murine LIF, and methods of producing LIF by the expression of the isolated DNA in suitable host cells. The invention further provides human and murine LIF polypeptides, pharmaceutical compositions containing LIF, and methods of use thereof.

Abstract: A novel peptide having molecular weight of 31,000 (SDS-PAGE) is obtained from aqueous extract of Natto or the culture of Bacillus natto by purification procedures including alcohol fractionation and/or salting out and hydrophobic chromatography, and the physicochemical properties, including the amino acid sequence, of the peptide are determined.The peptide is active in fibrinolysis, exhibits strong thrombolytic activity by oral administration and is useful as an oral thrombolytic agent.

Abstract: The present invention relates generally to a T cell growth factor. More particularly, the present invention relates to a T cell growth factor which comprises a glycoprotein which supports interleukin 2- and interleukin 4-independent growth of helper T cells especially from murine and human sources and further which is capable of augmenting proliferation of IL3- or IL4-responsive cells. Even more particularly, the present invention relates to the helper T cell growth factor P40, pharmaceutical compositions thereof, antibodies thereto and recombinant DNA clones thereof. The present invention also contemplates a method for inducing the proliferation of helper T cells as well as IL3- and Il4-responsive cells. The helper T cells growth factor contemplated herein is useful in the stimulation of specific cells in the immune system, either alone or in combination with IL3 or IL4.

Abstract: A vaccine effective against infection caused by group B Nesseria meningitidis microorganism is provided which comprises a purified protein antigenic complex weighing from 65 to 95 kDa, vesicles, and capsular polysaccharides. This vaccine is extracted from the cell membranes of the live microorganisms using detergent and enzyme. The method of producing an antimeningococcic hyperimmune gammaglobulin and the gammaglobulin produced by the method was provided. The gammaglobulin is obtained from vaccinees vaccinated with the vaccine.

Abstract: The present invention is directed to novel purified and isolated mite allergens possessing mite allergen activity with a molecular weight of about 94,000, about 40,000, about 16,000 or about 14,000 as determined by SDS-PAGE, which can be isolated from extracts of mite, and to a process for producing such mite allergens. The purified and isolated mites allergens of the present invention are useful as a pharmaceutical and a diagnostic composition for mite allergic diseases.

Abstract: Method for obtaining highly purified transferrin from a partially purified plasma fraction containing transferrin in which the starting fraction is concentrated. Its ionic strength is reduced and then transferrin is adsorbed onto a chromatographic column. Following elution, the transferrin can be further processed through to packaging in final containers. The final purified transferrin product is sterile, at least 95% pure and is substantially free of enveloped and non-enveloped viruses.

Abstract: Opossum whole serum exhibits a life saving property by neutralizing the lethality of venoms from all major families of poisonous snakes, and therefore an injection of Opossum serum can used as a novel treatment for many types of envenomation. Preferably, the injectable treatment for envenomation should be a composition obtained from the fraction of Opossum whole serum which contains the LTNF, i. e. the so called "LTNF-n", in purity. A method is given for the manufacture of a LTNF from the serum of an opossum (Didelphis virginiana) serum, by fractionating the opossum serum and isolating this select fraction from the plurality of fractions having an N terminal amino acid sequence given by SEQ ID No: 1. A short peptide was synthesized having SEQ ID No: 1. The synthetic peptide having the sequence SEQ ID No: 1 shows lethal toxin neutralizing activity similar to the natural LTNF from opossum or mongoose sera. The synthetic LTNF also has life saving utility.

Abstract: Purified .alpha..sub.1 -acid glycoprotein and a process for preparing purified .alpha..sub.1 -acid glycoprotein. The process comprises providing an impure protein fraction, binding contaminants, but not .alpha..sub.1 -acid glycoprotein, to a cation-exchange medium, and binding .alpha..sub.1 -acid glycoprotein to an anion-exchange medium, and eluting the .alpha..sub.1 -acid glycoprotein from the anion-exchange medium.

Abstract: In a method of producing a virus-safe biological preparation by heating while preserving a least 50% of its biologic activity, a biologially compatible tenside is added to the preparation before heating and heating is carried out in the presence of the same, whereupon the tenside, preferably, is eliminated.

Abstract: Rana pipiens eggs are subjected to mechanical processing in the presence of a weakly acidic buffer to produce an extract. The extract is subjected to ion-exchange chromatography and size-exclusion chromatography. Two pharmaceuticals resulting from these steps have activity against certain cancer cells. The amino acid sequences and compositions of two preferred embodiments are disclosed.

Abstract: A process 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: A purified, naturally derived melanogenic inhibitor protein capable of inhibiting melanogenesis in pigmentary cells has an amino acid sequence SEQ ID NO: 4. A method of producing melanogenic inhibitor protein comprises grafting mammalian skin onto a live host, permitting the mammalian skin to remain on the host for a predetermined period of time, removing the mammalian skin, and extracting the protein from the skin. Methods of controlling melanogenesis in pigmentary cells or selectively destroying melanoma cells comprise the steps of mixing an effective amount of melanogenic inhibitor protein, or an active segment, derivative, or analog thereof, with a suitable carrier, and applying this mixture to the pigmentary or melanoma cells.

Abstract: In this invention a novel antigen is provided for screening patients suspected of having celiac disease. The antigen is prepared by starting with human placenta tissue which is perfused with a herpes and collagenase buffer to obtain a signle cell suspension. This suspension is enriched to obtain an enriched protein portion and then separated out to obtain an embryonic celiac antigen (ECA). This ECA is used with serum from patients to effect binding of ECA with IgA in the serum while applying a human IgA antibody to the serum. The results are then read on a spectrophotometer to confirm or negate the presence of celiac disease.