Abstract: Disclosed is a process to remove tumour necrosis factor .alpha. (TNF.alpha.) or/and bacterial lipopolysaccharides (LPS) from an aqueous liquid, in particular blood, blood plasma or serum, in an extracorporeal perfusion system after removing corpuscular blood components if necessary, wherein(a) the pH value of the body fluid is adjusted to pH<6,(b) a precipitation reagent in the form of a polyanion is added,(c) precipitated substances are removed by filtration or/and centrifugation and(d) the resulting liquid is passed over an anion exchanger.

Abstract: A method for enriching rare cell populations from a whole blood sample by separating rare cell fractions from whole according to the relative charge density and/or the relative binding affinity for a leukocyte depletion solid phase matrix. The enrichment method may be operated stand alone, or as a pre or post-processing step in conjunction with a charge-flow separation method.

Abstract: Method for purifying an aqueous solution of raw albumin wherein contaminant proteins are bound by chromatography to a stationary solid phase, preferably a particulate phase, the purified albumin solution being collected as an effluent, in which a neutral phase charged with at least one compound chosen from the group formed by compounds containing C.sub.3 to C.sub.8 alkyl radicals and compounds containing sulfate groups is used as the stationary phase.

Abstract: Peritoneal fluid and peritoneal wash fluid provide a source of abundant apolipoprotein E. Methods for isolating apolipoprotein E from peritoneal fluid or peritoneal wash fluid are provided.

Abstract: A chromatography column packing material and column chromatographic methods using a silica gel based column packing material are disclosed. The silica gel material is coated by a phospholipid bilayer containing at least two lipids, at least one having an electric charge. Proteins are separated using this column packing material without denaturing because of the lipid coating. The protein molecule binds to the lipid bilayer coated silica gel and is then eluted by raising the temperature.

Abstract: A cellulose binding domain (CBD) having a high affinity for crystalline cellulose and chitin is disclosed, along with methods for the molecular cloning and recombinant production thereof. Fusion products comprising the CBD and a second protein are likewise described. A wide range of applications are contemplated for both the CBD and the fusion products, including drug delivery, affinity separations, and diagnostic techniques.

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

Abstract: Discoveries are disclosed that show particular aspects of recombinant DNA technology can be used successfully to produce hitherto unknown human keratinocyte growth factor (KGF) protein free of other polypeptides. These proteins can be produced in various functional forms from spontaneously secreting cells or from DNA segments introduced into cells. These forms variously enable biochemical and functional studies of this novel protein as well as production of antibodies. Means are described for determining the level of expression of genes for the KGF protein, for example, by measuring mRNA levels in cells or by measuring antigen secreted in extracellular or body fluids.

Abstract: Mocarhagin, a cobra venom protease, is disclosed. Pharmaceutical compositions and therapeutic uses of the protease are also provided.

Abstract: A novel homogeneous human cytokine, Natural Killer Stimulatory Factor, having the ability to induce the production of gamma interferon in vitro in human peripheral blood lymphocytes, and a pharmaceutical preparation containing it.

Abstract: A purified preparation of an anti-CDW52 antibody which exhibits on size exclusion chromatography: a single peak under non-reducing conditions and two major peaks under reducing conditions.The preparation preferably also exhibits on conventional SDS PAGE: one main band using a non-reduced sample and two main bands using a reduced sample.Additionally the preparation exhibits on reversed phase HPLC: a single sharp peak under non-reducing conditions and two major peaks under reducing conditions. Also a process of purifying an anti-CDW52 antibody, formulations containing such a purified preparation and uses thereof.

Abstract: A purified glycoprotein complex of over 1200 kD apparent native molecular weight having a sedimentation value of approximately 25S and having the ability to selectively bind human Mac-2 or interfere with PHA activation of lymphocytes, DNA sequences that encode the protein, and expression systems for expressing it, thus providing for medicaments that are useful for treating or diagnosing diseases, including cancer, infectious disease, and diseases of the immune system.

Abstract: A process for purifying an antibody is provided. In this process, a mixture containing the antibody and contaminant is subjected to low pH hydrophobic interaction chromatography (LPHIC) optionally at low salt concentration. The antibody is eluted from the column in the fraction which does not bind thereto. This process can be preceded and followed by other purification steps.

Abstract: A novel polypeptide with binding affinity for the p185.sup.HER2 receptor, designated heregulin-.alpha., has been identified and purified from cultured human cells. DNA sequences encoding additional heregulin polypeptides, designated heregulin-.alpha., heregulin-.beta.1, heregulin-.beta.2, heregulin-.beta.2-like, and heregulin-.beta.3, have been isolated, sequenced and expressed. Provided herein are nucleic acid sequences encoding the amino acid sequences of heregulins useful in the production of heregulins by recombinant means. Further provided are the amino acid sequences of heregulins and purification methods therefor. Heregulins and their antibodies are useful as therapeutic agents and in diagnostic methods.

Abstract: Suspension-polymerized, crosslinked copolymers of methacrylic anhydride yield beads suitable as precursors for weak-acid and other ion exchange resins, affinity chromatography materials and other materials which require spherical polymers having reactive anhydride groups for their preparation. Both gel and macroporous beads of these copolymers may be made by the process of the invention.

Abstract: A method is described for identifying, localizing and treating tumors in a patient. The method comprises assaying extracellular fluids, isolated from a patient, for the presence of elevated levels of ring shaped particles (RSP). The assay employs binding labeled RSP specific binding agent to the RSP.

Abstract: A method for processing a fatty acid-acylated protein, especially N-palmitoyl Lys.sup.B29 human insulin, to reduce the incidence of gelation by conducting such processing in the presence of a citrate buffering agent as the primary buffer which method allows such processing to be conducted at higher protein concentrations and with less temperature control than would otherwise be possible in the absence of the citrate buffer.

Abstract: Enzyme specific for bilirubin which has a phenol oxidase activity of less than 0.5% and an activity for biliverdin of less than 10%, a broad pH optimum as well as a good thermostability. The enzyme is obtainable from plants such as alfalfa and is suitable for the determination as well as for the degradation of bilirubin in particular in biological liquids.

Abstract: The present invention relates to shark cartilage extracts and to a method of producing the same, these extracts having anti-angiogenic properties (reduction of the area of blood vessels observed in vivo on experimentally induced tumors), tumor regressive activity in vivo as well as demonstrating a direct inhibitory effect on tumor cell lines. This process does not involve any denaturing solvent or product and does not involve the use of any enzymes. It consists of obtaining a blend of whole cartilage in an aqueous solution of neutral pH, preferably pure water, this blend being centrifuged and the pellet and supernatant kept for further processing. The pellet is lyophylized and tested for anti-tumor and anti-angiogenic activities in vivo and in vitro, with or without supernatant. The supernatant has been shown to have anti-angiogenic and tumor regressive activities in vivo. The composition of the supernatant has then been investigated by different ways.

Abstract: The process for producing the albumin preparation of the present invention comprises treating an aqueous albumin solution with an anion exchanger and a cation exchanger and subjecting the solution to heat treatment wherein a polyacrylamide type carrier having a cation exchange group is used as a cation exchanger. By using a polyacrylamide type carrier as a cation exchanger, the polymerization caused by the contaminating proteins can be suppressed, and a highly purified albumin preparation can be obtained.

Abstract: Highly purified factor XIII and methods of purifying factor XIII are disclosed. Factor XIII is purified from a biological fluid, such as a cell lysate. The methods provide factor XIII compositions that are greater than 99% pure with respect to contaminating proteins. The methods further provide factor XIII compositions wherein 1% or less of the factor XIII is activated factor XIII.

Abstract: There are disclosed (i) a purification process for obtaining a human BCDF having the intramolecular disulfide linkage and the stereostructure of natural type human BCDF which comprises subjecting to an oxidation reaction and a refolding treatment a reduced type human BCDF obtained by culturing a microorganism having a human BCDF gene integrated therein and solubilized with guanidine hydrochloride, characterized in that after the oxidation reaction, a gel filtration chromatographic treatment is conducted under the conditions of the guanidine hydrochloride concentration adjusted to 4-7M; (ii) a purification process for obtaining a natural type human BCDF monomer by removing the organic solvent from an organic solvent-containing solution of human BCDF, characterized in that the solution is passed through a gel filtration chromatographic column equilibrated with an organic solvent, followed by eluting according to a stepwise or linear gradient program; and (iii) a human BCDF purification process comprising an ion

Abstract: A method for preparing a macromolecular protein complex (MPC) from fibrinogen in human plasma by the steps of ammonium sulfate precipitation, dialysis and gel filtration is disclosed. The isolated MPC may be degraded by contacting with trypsin. The isolated MPC inhibited fibrinolysis induced with plasminogen but not with plasmin. Elimination of the MPC by means of chondroitin sulfate A restored normal fibrinolysis. An antibody to fibrin-binding peptides which are produced by trypsin degradation of MPC, was conjugated to plasmin. The anti-MPC peptide antibody/plasmin conjugate has the capacity to bind to MPC-rich thrombus and degrade it without activation of fibrin-bound plasminogen.

Abstract: The invention involves the purification of various glial mitogenic factors. In one aspect of the invention, vertebrate brain material is extracted, and the extract is then subjected to chromatographic separation. The glial mitogenic factors which can be obtained include a basic polypeptide with a molecular weight of from about 30 to about 36 kD, by reference to SDS-PAGE, and a second basic polypeptide factor which is from about 55 to about 63 kD as determined by SDS-page.

Abstract: A process for the production of biologically active recombinant neurotrophic factor from the NGF/BDNF family is described. The process is comprised of a) constructing a synthetic neurotrophic factor gene suitable for expression in a bacterial expression system; b) the synthetic neurotrophic factor gene is expressed in a bacterial expression system; c) the neurotrophic factor is solubilized and sulfonylated; d) sulfonylated neurotrophic factor is allowed to refold in the presence of polyethylene glycol and urea; and e) biologically active neurotrophic factor is isolated and purified.

Abstract: Disclosed is a process of separating protein using a polymeric composition. The composition includes a polymer formed from at least one monomer containing a polymerizable moiety chemically bonded to an anionic organic dye. The dye has an affinity for the protein to be separated. The process includes retaining the protein on the dye fraction and recovering the protein from the polymer.

Abstract: A vaccine effective against infection caused by Group B Neisseria meningitidis microorganism is provided which comprises a purified protein antigenic complex weighing from 65 to 95 kDa, vesicles, and capsular polysaccharide. This vaccine is extracted from the cell membranes of the live microorganisms using detergent and enzyme.

Abstract: The disclosure describes highly specific tryptase polyclonal antibodies, and a method to purify the antibodies. Specifically, the invention relates to polyclonal antibodies which have the capacity to capture tryptase out of solution, a process to generate the antibodies, and an enzyme-linked immunosorbent assay (ELISA) for human tryptase which utilizes the antibodies.

Abstract: The invention relates to novel polypeptides with antithrombin activity obtainable from extracts of tissues or secretions of leeches of the order Rhynchobdellida, particularly of the species Theromyzon tessulatum. The polypetides have molecular weights of about 14 kD, 9 kD and 3 kD and can be used in pharmaceutical compositions for the treatment of thrombosis related disorders and events.

Abstract: This invention relates to methods for the isolation and purification of the recombinantly expressed major protein component of chondroitinase ABC, which is referred to as "chondroitinase I", from Proteus vulgaris (P. vulgaris). This invention further relates to methods for the isolation and purification of the recombinantly expressed second protein component of chondroitinase ABC, which is referred to as "chondroitinase II", from P. vulgaris. These methods provide significantly higher yields and purity than those obtained by adapting for the recombinant enzymes the method previously used for isolating and purifying native chondroitinase I enzyme from P. vulgaris.

Abstract: This invention is intended to provide a process by which a complex formed by the interaction between one or more analytes to be measured and affinity substance can be more effectively separated from substances existing together therewith which tend to affect the detection of the complex, for example, free affinity substance, by using high pressure liquid chromatography (HPLC); and a process for measuring a trace component by utilizing said separating process.This invention is characterized in that a substance which has been modified with a substance capable of changing properties of the complex (a separation-improving substance) and has affinity for the complex is attached to the complex. Because of this characteristic, the invention is effective in that the position of elution of said complex in the HPLC can be freely controlled.

Abstract: The present invention provides an affinity separation method involving dynamic filtration.

Abstract: Described are methods for determining the potency of somatotropins. The level of biologically active bovine somatotropin protein in a bovine somatotropin sample is measured by size exclusion HPLC employing as the stationary phase a hydrophilic porous gel having an average particle diameter of about 5 .mu.m to about 15 .mu.m and the a mobile phase a buffered aqueous solution which is non-denaturing to the bovine somatotropin sample. The potency of the bovine somatotropin sample is determined based upon the level of biologically active bovine somatotropin protein so measured. In other embodiments, somatotropin is provided dissolved in a first buffer solution, and then chromatographed in a mobile phase comprised of a second buffer solution having a pH lower than the first buffer solution, to achieve an effective separation of biologically active protein from biologically-inactive large non-covalent soluble aggregates.

Abstract: A method is provided for isolating substantially intact cardiac troponin I from cardiac tissue comprising extracting the troponin I and purifying it in the presence of an effective mount of a mixture of protease inhibitors. The human cardiac troponin I, prepared by the present method is characterised by a molecular weight of about 28 kDa.

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 biologically active, stable protein with a molecular weight of 20,000 to 30,000 and an isoelectric point of 8.0-10.0 is produced from the culture fluid of Streptococcus pyogenes and purified. The purified biologically active protein produces lymphocyte proliferation, provides protection against bacterial and viral infection and restricts minor metastasis.

Abstract: A method of treating a patient having a panic disorder, the patient having an elevated CCK peptide plasma level, by lowering the plasma CCK peptide level of the patient. A further method provides a diagnosis of panic disorder in a patient by detecting if that patient's plasma contains elevated CCK peptide levels. A further method determines the efficacy of the drug for the treatment of panic disorder by detecting the ability of the drug to lower elevated CCK peptide levels in a model for panic disorder. Additionally, a method of dosing a patient having elevated CCK peptide levels with an antipanic disorder drug is characterized by administering the drug to a patient and monitoring the lowering of the elevated plasma CCK peptide levels of the patient.

Abstract: The present invention provides a composition of matter useful for enhancing the viability of hybridomas in culture which comprises a partially purified, cell-free extract derived from a medium conditioned by mitogen-stimulated macrophages, the extract being substantially free of the macrophage stimulating mitogen which had been added to the medium, and having within it a component characterized by the ability to enhance the viability of hybridomas in culture, and by an apparent molecular weight in the range from about 35,000 to about 45,000. Also provided are methods of preparing the composition of matter, and methods for enhancing the viability of hybridomas in culture.

Abstract: A protein which inhibits Epidermal Growth Factor-induced cellular proliferation is disclosed, also a method of producing the protein in virus-infected host cells, including purifying the protein such as by using C18 reverse phase HPLC. Additionally, therapeutic uses of the protein also are described.

Abstract: A preselected hemoglobin species is separated from contaminants having a different acidity from that of the preselected hemoglobin species, by an overload displacement chromatography process. To remove more acidic contaminants, the process is conducted under anion exchange conditions. To remove more basic contaminants, the process is conducted under cation exchange conditions. In either case, the exchange column is overloaded to displace the hemoglobin species therefrom with contaminants having greater affinity for the column, and using the impure hemoglobin solution as the displacer.

Abstract: A substantially homogeneous protein having cystic fibrosis transmembrane conductance regulator activity is provided. Also provided is a therapeutically effective composition for treating a subject having cystic fibrosis.

Abstract: This invention relates to methods of removing undesired antibodies from blood-derived compositions containing both the antibodies and coagulation factors, such that the coagulation factors are substantially retained in the composition. The undesired antibodies may be blood group antibodies. This invention also relates to compositions in which undesired antibodies have been removed and desired coagulation factors are retained. This invention further relates to methods of inactivating virus and removing undesired antibodies from blood-derived compositions containing virus, antibodies and coagulation factors without removing coagulation factors therefrom, and to the resulting compositions.

Abstract: A method for sequencing proteins on a polytetrafluoroethylene support is described. The support is preferably porous. The sample to be sequenced may be transferred directly, e.g., by blotting, to the support. Covalent binding of the sample to the support is not required.

Abstract: A method for correlating a peptide fragment mass spectrum with amino acid sequences derived from a database is provided. A peptide is analyzed by a tandem mass spectrometer to yield a peptide fragment mass spectrum. A protein sequence database or a nucleotide sequence database is used to predict one or more fragment spectra for comparison with the experimentally-derived fragment spectrum. In one embodiment, sub-sequences of the sequences found on the database which define a peptide having a mass substantially equal to the mass of the peptide analyzed by the tandem mass spectrometer are identified as candidate sequences. For each candidate sequence, a plurality of fragments of the sequence are identified and the masses and m/z ratios of the fragments are predicted and used to form a predicted mass spectrum.

Abstract: The present invention provides methods and reagents for sequencing amino acids. One embodiment of the method for determining the terminal amino acid of a polypeptide comprises the steps of (a) attaching (either covalently or non-covalently) the polypeptide to a solid support, (b) reacting the polypeptide with a compound described below, under conditions and for a time sufficient for coupling to occur between the terminal amino acid of the polypeptide and the compound, thereby yielding a polypeptide with a derivatized terminal amino acid, (c) washing the solid support to remove unbound material, (d) cleaving the derivatized terminal amino acid from the polypeptide with a cleaving agent, (e) ionizing the cleaved derivatized terminal amino acid, and (f) determining the molecular weight of the derivatized terminal amino acid, such that the terminal amino acid is determined.Within one embodiment, the compound is p-isothiocyanato phenethyl trimethylammonium and counterion salts thereof.

Abstract: Recovery of the 52/48 kDa tryptic fragment of vWF or peptide subfragments thereof produced in the form of inclusion bodies from recombinant host cells is carried out by providing a washed recombinant host cell suspension, adding a detergent and subjecting the cells to mechanical disruption. A second detergent is added, followed by another mechanical disruption and centrifugation to a pellet. The pellet is resuspended in a buffer and subjected to another mechanical disruption. Inclusion bodies are washed, resuspended and then recovered. In addition, endotoxins and DNA are removed from inclusion bodies containing the 52/48 kDa tryptic fragment of vWF or peptide subfragments thereof by mechanically disrupting the inclusion bodies in an aqueous buffer containing a detergent. A washed pellet is formed from these mechanically disrupted inclusion bodies and dissolved in a denaturant.

Abstract: A cDNA and a chromosomal DNA coding for the human B-cell differentiation factor were provided and the entire nucleotide sequence of said DNAs as well as the entire amino acid sequences of the polypeptide portion of mature human B-cell differentiation factor and the leader peptide were revealed. The method for producing human B-cell differentiation factor by a recombinant gene technology was also provided.

Abstract: A modified form of KL, the ligand for the c-Kit proto-oncogene, has been prepared wherein the protein is stabilized by an intermolecular covalent linkage. The protein can be prepared by expression of a recombinant protein which is dissolved in denaturant and refolded under conditions resulting in a disulfide linked dimer. Examples demonstrate the purification and characterization of this disulfide-linked cysteine dimer kit ligand (KL-CD) which contains at least one intermolecular disulfide bond and has at least ten-fold greater activity in promoting cell proliferation than native, non-covalently linked KL, as measured in in vitro assays.

Abstract: A process for separating small molecules from a sample containing small and large molecules using a porous separation medium having bimodal chemistry wherein the small molecules are first removed is disclosed.

Abstract: Lactoperoxidase, secretory component and lactoferrin are separated in high purity from milk and related materials such as whey with a single cation exchange resin. After adsorption on the cation exchange resin, elution is carried out with an aqueous solution having an ionic strength and pH selected to elute each separately. Lactoperoxidase is eluted first, secretory component second and lactoferrin last. Each is obtained in a purity of about 80% or greater. The cation exchange resin can be a cross-linked polysaccharide, cellulose or an acrylamide resin having carboxyl, sulfonic acid or phosphoric acid functional groups which may be attached with a spacer. The separated lactoperoxidase, secretory component and lactoferrin are biologically active and can be used in pharmaceuticals, cosmetics, foods, drinks and feeds.