Abstract: The present invention relates to methods useful for purifying materials using adsorbent chromatography, preferably in an expanded bed or packed bed configuration without the need to use a moving packed bed adapter, thereby improving elution characteristics for the sample molecule of interest.

Abstract: The present invention relates to cartilage extracts and to a method of producing the same. Shark cartilage extracts having anti-angiogenic, direct anti-tumor proliferating, anti-inflammatory and anti-collagenolytic activities have been obtained by an improved process. The process comprises the steps of obtaining a homogenate of cartilage in an aqueous solution, this homogenate being centrifuged and further fractionated to obtain a total extract having molecules of a molecular weight comprised between 0 to 500 KDa. The composition of the liquid extract has then been investigated by different ways. Further fractionation of this extract led to the preliminary characterization of some of its active components. Due to the multiplicity of biological activities of the total liquid extract, it can be used for treating numerous diseases or conditions such as those having components selected from the group consisting of tumor proliferation, angiogenesis, inflammation and collagenolysis.

Abstract: The inhibitors, obtainable from tissue or secretions of leeches typically of the order Rhynchobdellida, has the following terminal sequence: NH.sub.2 -Lys-Leu-Leu-Pro-Cys-Lys-Glu-Y-His-Gln-Gly-Ile-Pro-Asn-Pro-Arg- wherein Y represents any amino acid sequence; or a pharmaceutically acceptable salt, derivative or bioprecursor of said sequence, or an analogue or homologue thereof. Because of their extreme potency in the nanomolar range, they can be used to treat a number of diseases where protein cross-linking is important. They can be used for the treatment of Crohn's disease, tumor implantation, atherosclerosis, thrombotic microangiophathy, fibrous growths of the skin, acne, scar formation, membranous glomerulonephrits, cataracts, or infection with microfilarial nematodes. In particular, they can be used to reduce the stability of thrombi so that they are more susceptible to lysis by thrombolytic agents.

Abstract: The invention relates to a method for inducing bone formation via administration of an extract of devitalized, freeze dried Saos-2 cells. The extract is prepared by contacting the freeze dried Saos-2 cells, which are devitalized in the act of freeze drying, with a weak denaturing agent of the type used to separate proteins from cells. The extract does have properties superior to comparable treatment using whole Saos-2 cells. The extract is then further treated by applying it to a removing separating gel, and then fractions from the gel which have a molecular weight of from about 10 kd to about 60 kd. Also described are the extract itself, and formulations containing it.

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. Optionally, the methods further comprise administering antigen presenting cells sensitized with complexes of hsps 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. In a specific embodiment, the effective amounts of the complex are in the range of 0.1 to 9.

Abstract: The present invention concerns the purification of keratinocyte growth factors.

Abstract: The invention relates to a method of purifying cholera toxin using a matrix with at least one ion chosen from among matrix with Ni.sup.+2, Co.sup.+2, Cd.sup.2 or Zn.sup.+2 immobilized thereon. It is possible thereby to selectively elute the B subunit for cholera toxin from the matrix.

Abstract: The invention relates to a method for inducing bone formation via administration of an extract of devitalized, freeze dried Saos-2 cells. The extract is prepared by contacting the freeze dried Saos-2 cells, which are devitalized in the act of freeze drying, with a weak denaturing agent of the type used to separate proteins from cells. It is not clear that the active ingredient in the extract is a protein; however, the extract does have properties superior to comparable treatment using whole Saos-2 cells. Also, described are the extracts themselves and various compositions containing it.

Abstract: Methods are provided for large scale purification of neurotrophins, including mature NGF, suitable for clinical use. The methods provide means to separate neurotrophins from various less desirable misprocessed, misfolded, size, glycosylated, or charge forms. Compositions of neurotrophins, including mature NGF, substantially free of these variants are also provided.

Abstract: The present invention relates to a process for purifying recombinant coagulation factor VIII by loading an aqueous solution containing factor VIII onto a hydrophobic interaction chromatography (HIC) gel, wherein at least one surfactant is present in the aqueous solution and/or a buffer solution used to equilibrate the gel prior to loading the aqueous solution onto the gel. The presence of a surfactant when loading the solution containing factor VIII onto the HIC gel, makes it possible to efficiently separate the intact and active factor VIII molecules from molecules with structural deviations. With the present invention it is further possible to reduce the content of DNA and/or CHO cell contaminants considerably and increase the activity of the factor VIII product to a hitherto unknown extent.

Abstract: A method of separating albumin from serum by ion exchange chromatography with membrane adsorbers characterized by high productivity and yields of high purity albumin. The separation of the albumin is carried out on highly basic anion exchange membranes and on highly acidic cation exchange membranes. The albumin fraction can be eluted from the anion exchange membrane such that it can be fed directly to the cation exchange membrane without any special conditioning and the albumin can be extracted therefrom as an end product.

Abstract: One aspect of the present invention is a purified human GMP synthetase and a method of purifying it from naturally occurring sources. Another aspect of the present invention is a recombinant human GMP synthetase as well as DNA sequences coding for human GMP synthetase, expression vectors comprising such a coding sequence, and host cells transformed with these expression vectors capable of producing human GMP synthetase. Also forming part of this invention is a recombinant process for the production of GMP synthetase. Further provided is a method of purifying GMP synthetase from natural or recombinant sources. Other aspects of the invention include antibodies to human GMP synthetase and the use of such antibodies to assay for human GMP synthetase. Another aspect of this invention is the use of purified naturally occurring human GMP synthetase or recombinant human GMP synthetase to identify inhibitors of GMP synthetase activity.

Abstract: Methods are herein provided for the isolation and purification of zeamatin, an antifungal protein from corn. The subject methods use capture chromatography and reverse phase chromatography. The methods herein described is superior to prior art techniques as it the eliminates ammonium sulfate precipitation and centrifugation steps.

Abstract: A glioblastoma-derived angiogenesis inhibiting factor is described. The material is induced in presence of wild type p53, but not by several mutated forms of p53. Various uses of the material are described.

Abstract: The instant invention relate to a new method for the purification of proteins using copper chelate-affinity chromatography, wherein the impure or pre-purified protein is adsorbed on immobilized copper(II) ions, optionally washed with buffer and deionized water, washed with a solution of a Lewis-base, and finally eluted with deionized water.

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 lyophilized 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: A scalable method for the production of highly purified plasmid DNA in Escherichia coli is described, which method includes growing plasmid-containing cells to a high biomass in exponential growth and lysing the cells by raising the pH of the culture to a carefully controlled pH value in which chromosomal DNA is denatured but plasmid DNA is reversibly renatured. The method has been developed for the production of pharmaceutical grade DNA for use in in vivo and ex vivo gene therapy.

Abstract: The present invention is directed to a simple and efficient process for the recovery of a biologically active glycoprotein from a biological fluid containing it.In a preferred embodiment, it comprises:a) contacting a filtered culture fluid with a dihydroxyboronyl bearing chromatographic supportb) eluting with a first eluting buffer and contacting this eluate directly with an anion exchange matrix bearing quaternary ammonium functional groupsc) eluting with a second eluting buffer and collecting the product therefrom.

Abstract: Disclosed is a 12-kDa PPD protein isolated and purified from the purified protein derivative, the major fraction of Mycobacterium tuberculosis that protects mice against the induction of experimental autoimmune encephalomyelitis (EAE), and salts, functional derivatives, analogs and active fractions thereof, and pharmaceutical compositions comprising them for the treatment of autoimmune diseases.

Abstract: The invention relates to a process for the preparation of PDGF-A and to biologically active PDGF-AA and PDGF-AB.

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: Functionalized silicas which can be used as core supports for a broad variety of chiral stationary phases may be conceptually represented as T--O--Si--U. T represents a refractory inorganic oxide, and T--O--Si arises from reaction of an organosilane with the surface hydroxyl groups of the refractory inorganic oxide. U represents a polyamine, related to poly(ethyleneamines), tris(2-aminoethyl) amine, or alkyleneoxyamines of glycerine.

Abstract: The present invention provides antiviral proteins, peptides and conjugates, as well as methods of obtaining these agents. The antiviral proteins, peptides and conjugates of the present invention can be used alone or in combination with other antiviral agents in compositions, such as pharmaceutical compositions, to inhibit the infectivity, replication and cytopathic effects of a virus, such as a retrovirus, in particular a human immunodeficiency virus, specifically HIV-1 or HIV-2, in the treatment or prevention of viral infection.

Abstract: A new class of proteins and methods related thereto are presented. The proteins, which can be characterized as catalysts of the extension of plant cell walls and the weakening of the hydrogen bonds in pure cellulose, are referred to as expansins. Two proteins have been isolated by fractionation techniques from washed wall fragments of cucumber hypocotyls, referred to as "cucumber expansin-29" and "cucumber expansin-30" (abbreviated cEx-29 and cEx-30, with respect to their apparent relative masses as determined by SDS-PAGE). Moreover, three peptide fragments from the purified cEx-29 protein were sequenced, then oligonucleotide primers were designed to amplify a portion of the expansin cDNA using polymerase chain reaction with a cDNA template derived from cucumber seedlings, and then the PCR fragment was used to screen a cDNA library to identify full length clones.

Abstract: A method and vaccine for treatment of pythiosis in humans and animals is described. In particular a vaccine comprising a mixture of extracellular and intracellular proteins is described. The vaccine enables cures of chronic pythiosis in some patients.

Abstract: A method for enriching rare cell populations from a whole blood sample by separating rare cell fractions from whole blood sample according to the relative charge density and/or the relative binding affinity for a leukocyte depletion solid phase matrix is described. 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: A method for purifying protein containing histidine residues using immobilized metal, affinity chromatography. The hydrophilic index of the histidine residues is determined (HI). If the HI is at least 2 the pH of the solution containing the protein is adjusted to about 6.75 to 7.2 and applied to the IMAC column such that the protein binds to the column.

Abstract: Porous polymer monoliths are made thermally responsive by functionalizing/grafting the pores with thermally responsive polymers and copolymers. Depending on the reaction conditions employed, the grafted polymer can either completely block flow through micrometer-sized pores in the monoliths or control the flow rate through the monoliths. The grafted monoliths are useful as thermal gates, thermal valves, and for isocratic hydrophobic interaction chromatography of proteins.

Abstract: A method of preparing a novel, sterile, receptor rich-albumin molecule which utilizes the disinfecting properties of iodine by reacting an iodine donating material or solution with a pure preparation of albumin, and preferably subsequently removing the iodine. The resulting iodine has improved binding properties because the production method strips bacterial endotoxin and other previously bound substances from the albumin. The improved binding site capacity of the albumin product is advantageously used as an adjunct in removing toxins by means of exchange transfusions. Because iodine disinfects the albumin typical pasteurization and related additives are unnecessary.

Abstract: A final drug product comprises a plasma protein selected from the group consisting of coagulation factor VIII and factor IX, in an aqueous solution. The concentration of oxygen in the solution is reduced and/or the solution contains an antioxidant. The solution further contains a carbohydrate in a concentration of at least 350 mg/ml. The protein activity after storage for at least 6 months at a temperature of from 0.degree. C. to 40.degree. C. is from 70% to 130% of its initial value. In a process for preparing the final drug product and a method for improving the long-term stability of coagulation factor VIII or factor IX in an aqueous solution, a carbohydrate is included in the solution in a concentration of at least 350 mg/ml and the solution is stored in its final container under an oxygen-reduced atmosphere or includes an antioxidant.

Abstract: The invention relates to a method for purification of a mixture of hydroxamate derivatized protein/proteins and native protein which is characterized by treating the mixture with immobilized metal and thereby adsorbing the hydroxamate derivatized protein/proteins on the immobilized-metal and recovering the native protein. The protein could be IGF-L. It also relates to a process for the production of a native protein which is characterized by expression of the protein as a fusion protein, cleavage of the fusion protein by hydroxylanine, separation of native protein from hydroxamate derivatized protein by adsorbing the hydroxamate derivatized protein on immobilized metal and directly recovering the native protein. The use of immobilized metal affinity chromatography for separation of native protein from hydroxamate derivatized protein is also claimed.

Abstract: The present invention relates to processes for the purification of proteins. More specifically, methods for solubilizing and purifying proteins expressed in an insoluble form using low concentrations of chaotropic agents, such as guanidine salts, are provided. Also provided are methods for refolding proteins solubilized according to the present invention.

Abstract: This invention relates generally to modified porous solid chromatographic media and processes for the preparation and use of same. In particular, chromatographic media of porous mineral oxide, polymeric, or polymer-coated mineral oxide supports are disclosed which are characterized by a reversible high sorptive capacity and high intraparticle mass transfer rates. In order to prevent non-specific adsorption of or interaction with biomolecules, these supports may be passivated by use of a passivation mixture comprising a main monomer, a passivating monomer, and a crosslinking agent, which mixture upon polymerization results in substantial elimination of the undesirable non-specific interaction with biomolecules.

Abstract: A process for preparing thrombin which comprises treating a mixture comprising prothrombin, factor Xa, factor Va, and phospholipids with calcium ions, at a pH of 6.0-7.0 is provided. In particular the pH of 6.0-7.0 may be generated by the addition of the calcium ions or by buffering the preparation to a pH of 6.0-7.0. Thrombin preparations so produced may be subjected to further purification and are particularly stable even when substantially free of exogenous stabilizing agents such as proteins, sugars, polyol and mixtures thereof, and may be subject to freeze-drying and a virus inactivation by heat treatment.

Abstract: A method for preparing and isolating a transformation vector containing CSF/cDNA is described. The method comprises:preparing RNA from a cell that produces CSF;preparing polyadenylated messenger RNA from said RNA;preparing single stranded cDNA from said messenger RNA;converting the single stranded cDNA to double stranded cDNA;inserting the double stranded cDNA into transformation vectors and transforming bacteria with said vector to form colonies;picking pools of 200 to 500 colonies each and isolating plasmid DNA from each pool;transfecting the plasmid DNA into suitable host cells for expressing CSF protein;culturing the transfected cells and assaying the supernatant for CSF activity; andselecting CSF positive pools and screening the colonies used to make the pool to identify a colony having CSF activity. Also described are a cDNA coding for a protein having CSF activity (i.e.

Abstract: Embodiments of the present invention relate to methods and articles of manufacture for the detection of Giardia cysts and Crytosporidium oocysts.

Abstract: The invention provides a nucleic acid encoding a human cytochrome P450 2E1 comprising a 5' terminal deletion of 63 nucleotides, thereby encoding methionine at the first codon position of the 5' terminus, the codon ATG in the second codon position of the 5' terminus, and silent adenine and thymine nucleotide substitutions in the 5' region. The invention also provides a nucleic acid encoding a human cytochrome P450 2C10 comprising a 5' terminal deletion of nucleotides 7 through 60, thereby encoding methionine at the first codon position and alanine at the second position of the 5' terminus, and silent adenine and thymine nucleotide substitutions in the 5' region. The present invention also provides a method of purifying a recombinant cytochrome P450 protein from a host cell culture comprising the steps of: a. fractionating the host cells to prepare their membranes; b. adding a non-ionic detergent in a concentration of between 0.

Abstract: A method for purifying a plant protein comprises: (a) preparing a crude plant extract; (b) passing the crude plant extract with a first buffer solution through a guard column comprising a hydrophobic interaction chromatography medium of hydrophobicity sufficiently high to prevent tannins and polyphenols from eluting from the column in the presence of the buffer solution, but sufficiently low that a protein fraction elutes with the first buffer solution; (c) passing the protein fraction through a capture column coupled in series to the guard column, the capture column comprising a hydrophobic interaction chromatography medium of hydrophobicity sufficiently high to prevent the protein from eluting in the presence of the buffer solution; and (d) eluting the protein from the capture column as a purified fraction. Preferably the plant is miracle fruit, the protein is Miraculin, and the method comprises ion exchange chromatography and gel filtration chromatography of the purified fraction of the protein.

Abstract: A fimbrial agglutinogen preparation is prepared from a bordetella strain, particularly a B. pertussis strain, by a multiple step procedure involving extraction of the fimbrial agglutinogens from cell paste and concentrating and purifying the extracted material. The fimbrial agglutinogen preparation may be used to prepare acellular pertussis vaccines with other pertussis antigens, including pertussis toxin or toxoid thereof, the 69 kDa protein and filamentous hemagglutinin and other Bordetella antigens.

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: 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: The present invention comprises an aggregate endothelial inhibitor and method of use therefor. The aggregate endothelial inhibitor comprises a protein having a molecular weight of between approximately 45 kilodaltons and 65 kilodaltons as determined by reducing polyacrylamide gel electrophoresis, and having an amino acid sequence substantially similar to that of a murine plasminogen fragment beginning at amino acid number 98 of a murine plasminogen molecule.

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: 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 amount 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: 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: The present invention provides methods for separating ligands from binding proteins. The methods include acidic separation and size separation. The methods of the present invention are particularly useful for separating insulin-like growth factors from insulin-like growth factor binding proteins.

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: The invention relates to chemotactic factors for human spermatozoa that are purifiable from human follicular fluid. The factors are of peptidic and hydrophilic nature and have, one, a molecular size of about 13 kDa and, the other, an apparent molecular size smaller than 1.3 kDa, as determined by high pressure gel filtration. They are for use in procedures related to human fertilization, such as in various types of assisted fertilization, particularly artificial insemination, in vitro fertilization, micromanipulation and direct microinjection of sperm into oocytes.

Abstract: A process for extraction of myelin basic protein from myelin containing tissue, such as central nervous system tissue, which process comprises the following steps:extraction of the myelin basic protein from myelin containing tissue with an organic solvent selected from the group consisting of chloroform and compounds having a polarity similar to that of chloroform;incubation of the organic phase in the presence of a lower aliphatic alcohol or propylene glycol;transfer of the myelin basic protein from the lower aliphatic alcohol/organic solvent mixture to an aqueous phase with the aid of hydrogen ions (protons); andrecovery of the purified myelin basic protein. The invention also relates to the product obtainable by the process.

Abstract: A composition is disclosed which comprises a solution of human serum albumin essentially free of chemicals used in processing. The preparation is also essentially free of metals such as aluminum. The composition is 100% pure by cellulose acetate electrophoresis and is essentially monomeric when tested by high pressure liquid chromatography. The turbidity is less than 5 N.T.U. (National Turbidity Units). This preparation has a substantially longer shelf life and remains biologically active longer than products currently available. The novelty of this product is also such that it does not leach metallic substances such as aluminum from its closure. Novel applications of process methodology are taught in the preparation of this composition and a novel preparation results from essentially non hemoglobin containing albumin sources such as Source Plasma (Human).