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, using a readily available sulfated chitin as an adsorbent, can permit platelet factor-4 to be recovered through specific adsorption from a solution containing the same factor, in by far increased yields as compared with the conventional process utilizing a heparin-immobilized affinity column, and provides the process for isolating, through purification platelet factor-4 which is suited for a commercial-scale, mass production process, wherein there can be offered the advantages of utilization of more readily available sulfated chitin, simplified procedure and improved production yields for the objective substance.

Abstract: The invention relates to substantially pure transfer factor with a specific activity of at least 5000 units per absorbance unit at 214 nm. The present invention also relates to a process for preparing the transfer factor from cell lysates. The present invention includes the use of substantially pure transfer factor with a specific activity of at least 5000 units per absorbance unit at 214 nm to treat infectious diseases.

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 methods and compositions for eliciting an immune response and the prevention and treatment of primary and metastatic neoplastic diseases and infectious diseases. The methods of the invention comprise administering a composition comprising an effective amount of a complex, in which the complex consists essentially of a heat shock protein (hsp) noncovalently bound to an antigenic molecule. "Antigenic molecule" as used herein refers to the peptides with which the hsps are endogenously associated in vivo as well as exogenous antigens/immunogens (i.e., with which the hsps are not complexed in vivo) or antigenic/immunogenic fragments and derivatives thereof. In a preferred embodiment, the complex is autologous to the individual. The effective amounts of the complex are in the range of 10-600 micrograms for complexes comprising hsp70, 50-1000 micrograms for hsp90, and 10-600 micrograms for gp96.

Abstract: Disclosed are chromatography methods and matrix geometries which permit high resolution, high productivity separation of mixtures of solutes, particularly biological materials. The method involves passing fluids through specially designed chromatography matrices at high flow rates. The matrices define first and second interconnected sets of pores and a high surface area for solute interaction in fluid communication with the members of the second set of pores. The first and second sets of pores are embodied, for example, as the interstices among particles and throughpores within the particles. The pores are dimensioned such that, at achievable high fluid flow rates, convective flow occurs in both pore sets, and the convective flow rate exceeds the rate of solute diffusion in the second pore set. This approach couples convective and diffusive mass transport to and from the active surface and permits increases in fluid velocity without the normally expected bandspreading.

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 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: 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 purified protein having a molecular weight of 40 kD by SDS-PAGE that produces firefly luciferin when combined with D-cysteine and firefly oxyluciferin and isolated from firefly species is provided, as well as methods of making and using the protein for the continuous regeneration of of firefly luciferin.

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: 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 present invention is directed to methods of detecting angiostatin protein which is an endothelial cell inhibitor. The angiostatin protein is a protein isolated from blood or urine that is eluted as single peak from C4-reverse phase high performance liquid chromatography. One method of the present invention includes combining a sample suspected of containing angiostatin protein with an antibody which is specific for angiostatin protein. Another method of the present invention includes isolating the protein and detecting the presence of angiostatin protein by performing an endothelial cell proliferation inhibiting assay.

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 process for the production of a solubilized protein is disclosed, which is obtained by reducing disulfide bonds in a disulfide bond-containing water-insoluble protein material into mercapto groups and subsequently converting a part or an entire portion thereof into carboxymethyldisulfide groups. In particular, the disclosed process for the production of a solubilized protein which comprises (a) treating a disulfide bond-containing water-insoluble protein material with an aqueous alkaline solution of a reducing agent, and (b) reacting the protein treated by step (a) with thioglycolic acid in the presence of an oxidizing agent under a weakly acidic to a weakly alkaline condition. A process for the production of regenerated protein products, which comprises regenerating disulfide bonds in the solubilized protein, is also disclosed.

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: Disclosed are specifically designed binding, "protein imaged" sorbents which reversibly bind with high specificity and affinity a preselected macromolecule, specifically a protein. The sorbents define one or more cavities which have a binding surface complementary in shape to the molecular surface of the macromolecule and a plurality of positively and negatively charged chemical moieties spatially distributed in a mirror image and charge inverse of a subset of the ionizable groups on the molecular surface of the macromolecule. Also disclosed are methods of producing such sorbents, useful over a range of conditions, for both preparative and analytical chromatographic separations or for use in various types of analyses.

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: The present invention permits human urinary thrombomodulin to be purified by a simple and practical procedure, while it enables even a starting material with a lowered content of thrombomodulin to be purified by greater purification rate than the one with a high content of thrombomodulin.

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: 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 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: 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: A method is provided for determination of the biological activity, purity and/or homogeneity of recombinant human growth hormone ("rhGH") using an analytical chemistry test. The method is based on distribution of the rhGH between two or more immiscible aqueous phases and the subsequent determination of the ratio of rhGH concentration in the phases. The ratio is then used as a relative measure of biological activity, homogeneity, and purity, when calibrated against a sample of rhGH that was previously characterized with respect to its activity, homogeneity and purity. Typical applications of this test include quality control, quality assurance, biological identity testing, etc.

Abstract: A composition comprising coagulation factor VIII and a non-ionic surfactant such as block copolymers, e.g., polyoxamers or polyoxyethylene (20) sorbitan fatty acid esters, e.g., polysorbate 20 or polysorbate 80 as stabilizer is provided. The composition can also comprise sodium chloride, calcium chloride, L-histidine and/or sugars or sugar alcohols.

Abstract: This invention provides a selective cellular fibronectin (cFN) adsorbent utilizing a nonwoven cellulose sulfate fabric and a method for fractional purification of FN which-comprises contacting an FN matter containing both plasma fibronectin (pFN) and cellular fibronectin (cFN) with the nonwoven cellulose sulfate fabric to separate pFN and cFN from each other. By the fractional purification method of the invention, cFN and pFN can be fractionated in an expedient manner and with high efficiency and both pFN and cFN can be recovered in high purity and good yield.

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 method for renaturing a denatured protein is provided comprising contacting a solution or suspension of said denatured protein in a detergent-free liquid medium with an amount of a cyclodextrin effective to renature said protein, and optionally, recovering the renatured protein in essentially pure form.

Abstract: The present invention is directed to a family of phospholipid binding and transporting proteins, a method for preparing same from fungi, and their use in cosmeticology, the agri-foodstuffs industry and pharmacology Phospholipid proteins are capable of binding, transporting and/or rearranging lipids between membranes, optionally in combination with active principles. Furthermore, the phospholipid proteins are hydrophobic and acidic, have a molecular weight of under 50 kDa, and may be prepared from a non-toxic filamentous fungus capable of developing on a lipid-enriched medium, particularly from raw extracts of fungi.

Abstract: Provided by the present invention are novel methods of factor IX protein recovery and purification.

Abstract: Compositions enriched for Neutrophil Inhibitory Factor which inhibit neutrophil activity including adhesion to vascular endothelial cells are provided. Such compositions may comprise a glycoprotein isolated from nematodes, particularly of the genus Ancylostoma. These compositions are useful in the therapy of conditions which involve abnormal or undesired inflammatory responses.

Abstract: The present invention describes a method for controlled activation and degradation of FVII during purification whereby a solution of Factor VII is subjected to a number of chromatographic purification steps wherein Zn.sup.++ is present in at least one of the purification steps.The present invention also describes a method for controlled activation and degradation of FVII wherein a solution of FVII is applied to a number of anion exchange and immunoaffinity columns.

Abstract: Disclosed herein is a process for purifying hirudin from a solution containing hirudin and other substances, characterized in that the process comprises a step of subjecting the solution to a metal ion affinity chromatography wherein copper ion (Cu.sup.++) is used as a metal ion and a phosphate buffer is used as an eluent.

Abstract: A method for purifying recombinant HIV gp120 so as to provide a glycopeptide having protein/protein binding properties substantially identical to natural viral HIV gp120, which comprises fractionating a composition containing crude gp120 sequentially using (1) ion exchange chromatography, (2) hydrophobic-interaction chromatography, and (3) size exclusion filtration, collecting at each step a fraction that exhibits specific binding affinity for CD4 peptide. The process is carried out in the absence of any affinity purification steps or any steps (such as reverse-phase HPLC) that use contact protein with organic solvents. The product obtained by this method is a purified, full-length, non-fusion recombinant HIV gp120 glycoprotein having protein/protein interaction properties substantially identical to gp120 as presented on an HIV virus, including binding affinity for CD4 and binding affinity for at least one antibody capable of neutralizing HIV infectivity.

Abstract: The present invention relates to an isolated protein or polypeptide capable of production by T cells and capable of ultimately activating production of B cells. The present invention further provides an isolated DNA molecule encoding the protein or polypeptide capable of production by T cells and capable of ultimately activating production of B cells. The isolated DNA molecule can be inserted as a heterologous DNA in an expression vector forming a recombinant DNA expression system for producing the protein or polypeptide. Likewise, the heterologous DNA can be incorporated in a cell to achieve this objective. The isolated protein or polypeptide of the present invention can be combined with a pharmaceutically-acceptable carrier or used alone for administration to mammals for activating production of B cells.

Abstract: A method and apparatus for carrying out affinity purification of a ligate. The method comprising, (a) providing a ligate containing liquid to a first side of at least one porous hollow fiber membrane with a ligand immobilized thereto that binds and separates the ligate from the liquid, (b) withdrawing a first portion of the liquid from the first side of the porous hollow fiber membrane, (c) recirculating the first portion of liquid to the first side of the porous hollow fiber membrane, (d) repeating steps (a) to (c) until a majority of the liquid has flowed through the porous hollow fiber membrane, and (e) providing an elution solution to one side of the porous hollow fiber membrane under a pressure sufficient to cause the elution solution to flow into and through the membrane to effect disassociation of any ligate-ligand bonds wherein any ligate bound to the ligand is eluted with the elution solution.

Abstract: Disclosed is a method for purifying Human interleukin-5 (IL-5) a single chromatographic step. After removing cells from a cell culture expressing human interleukin-5, IL-5 is purified by first adjusting the culture supernatant to the calculated pI value of mature IL-5 (pI=7.44) and then passing the conditioned supernatant through tandem linked anion- and cation-exchange columns. The resulting pass-through fraction contains the IL-5 and is devoid of all other contaminating proteins. An optional hydrophobic-interaction chromatography step is disclosed for positive selection of IL-5 and in order to concentrate the preparation. Pure IL-5 was recovered with a high overall yield (>90%), was N-glycosylated and was entirely homodimeric.

Abstract: The invention relates to a process for preparing a concentrate of Factor VIII-von Willebrand factor complex having high specific activity from total (non-cryoprecipitated) plasma.The process comprises pre-purifying by means of a double treatment with barium chloride and with aluminium hydroxide.The process then comprises purification by chromatography on an anion exchange resin, of the DEAE-Fractogel type.The process includes a step of viral inactivation by means of a treatment with solvent-detergent.The process also makes it possible to recover fibrinogen, albumin, immunoglobulins, antithrombin III, fibronectin and prothrombin complex, from the same plasma.The different concentrates obtained using the process according to the invention are intended, in particular, for therapeutic use.

Abstract: Polyolefin particles are chemically modified by oxidation to provide a large surface area and high loading. The particles result in low back pressure in column systems, and are economical to manufacture. The particles are useful as supports in a wide range of applications including general organic as well as biopolymer synthesis, library methods, purification processes and enzyme mediated processes. In a preferred embodiment, polyethylene or polypropylene particles are oxidized in a solution containing trifluoroacetic acid or trifluoromethane sulfonic acid, chromium trioxide and sulfuric acid to provide the particles with a chemically reactive irregular surface and open channels that extend below the surface and up to essentially the length of the radius of the particles resulting in increased surface area and decreased density. The particles have pendant functional groups produced by the oxidation and/or by subsequent chemical reaction.

Abstract: Gelled material compositions for use as adsorbents and ion exchangers. The composition including a specific extractant, an organic solvent and a halopolymer modified by substitution with a radical of a substance compatible with the specific extractant. Optionally the composition may be in form of beads.

Abstract: In order to remove tumor necrosis factor .alpha. (TNF.alpha.) or/and bacterial lipopolysaccharides (LPS, endotoxin) extracorporeally from whole blood or/and blood plasma in an extracorporeal perfusion system, the blood or plasma is passed over a cation exchanger and an anion exchanger material. A device according to the invention for the extracorporeal treatment of patient's blood or plasma therefore contains a cation exchanger material and an anion exchanger material wherein these materials are contained in at least one compartment of an extracorporeal perfusion system.