Abstract: The present invention provides methods for quantitation of glycated protein in a biological sample using a solid support matrix by making a first bound protein measurement total bound protein under conditions where both glycated and non-glycated protein bind to the support in making a second bound protein measurement under conditions where glycated protein is bound to the support and non-glycated protein is not substantially bound. Diagnostic devices and kits comprising the methods of the present invention are also provided.

Abstract: The present invention provides methods for determining a ratio of an amount of a glycated form of a protein to a total amount of the protein in a sample containing the glycated protein, the glycosylated protein, or the glycoprotein. The method incorporates lateral flow test strip or vertical flow test strip devices having negatively charged carboxyl or carboxylate groups and hydroxyboryl groups immobilized and interspersed on a solid support matrix. The solid support matrix may include derivatives of cellulose (e.g., carboxy cellulose) derivatized with carboxylic acid (e.g., carboxylate, carboxyl) groups and hydroxyboryl compounds including phenylboronic acid (e.g., phenylborate), aminophenylboronic acid, boric acid (e.g., borate), or other boronic acid (e.g., boronate) compounds. The present invention is usefi.il for monitoring glycation or glycosylation of hemoglobin or albumin for monitoring glycemic control (e.g., glycemia in diabetes).

Abstract: This invention provides methods of retentate chromatography for resolving analytes in a sample. The methods involve adsorbing the analytes to a substrate under a plurality of different selectivity conditions, and detecting the analytes retained on the substrate by desorption spectrometry. The methods are useful in biology and medicine, including clinical diagnostics and drug discovery.

Abstract: The invention provides methods and compositions for azide tagging of biomolecules. In one embodiment of the invention, proteins are tagged by metabolic incorporation of prenylated azido-analog substrates. Examples of such analogs are azido farnesyl diphosphate and azido farnesyl alcohol. The azido moiety in the resulting modified proteins provides an affinity tag, which can be chemoselectively captured by an azide-specific conjugation reaction, such as the Staudinger reaction, using a phosphine capture reagent. When the capture agent is biotinylated, the resulting conjugates can be detected and affinity-purified by streptavidin-linked- HRP and streptavidin-conjugated agarose beads, respectively. The invention allows detection and isolation of proteins with high yield, high specificity, and low contamination without harsh treatment of proteins.

Abstract: A process for preparing lysozyme is provided which is characterized by mixing the egg white or its diluted solution with diatomaceous earth, kaolin, zeolite, or the mixtures thereof, and followed by eluting the adsorbed lysozyme with a salt solution. The diatomaceous earth, kaolin, and zeolite can specifically adsorb the lysozyme in egg white, and the adsorbed lysozyme can be easily eluted with a salt solution. According to the process of the present invention, the lysozyme in egg white can be easily and effectively isolated.

Abstract: The invention relates to a process for synthesis, by inverse bead polymerization of a monomer phase, of a bead-like, cross-linked, hydrophilic copolymer which has binding activity toward ligands containing nucleophilic groups. The invention relates to support polymer materials with high binding capacity for penicillin acylase and low swelling factor, as well as to use of the same.

Abstract: Disclosed are methods and compositions for the identification, characterization and inhibition of farnesyl protein transferases, enzymes involved in the farnesylation of various cellular proteins, including cancer related ras proteins such as p21ras. One farnesyl protein transferase which is disclosed herein exhibits a molecular weight of between about 70,000 and about 100,000 upon gel exclusion chromatography. The enzyme appears to comprise one or two subunits of approximately 50 kDa each. Methods are disclosed for assay and purification of the enzyme, as well as procedures for using the purified enzyme in screening protocols for the identification of possible anticancer agents which inhibit the enzyme and thereby prevent expression of proteins such as p21ras. Also disclosed is a families of compounds which act either as false substrates for the enzyme or as pure inhibitors and can therefore be employed for inhibition of the enzyme.

Abstract: Multifunctional, polyionic copolymers with molecular architectures and properties optimized for specific applications are synthesized on/or applied to substrate surfaces for analytical and sensing purposes. The coatings are particularly useful for suppression of non-specific interaction, adsorption or attachment of molecular or ionic components present in an analyte solution. Chemical, biochemical or biological groups that are able to recognize, interact with and bind specifically to target molecules in the material containing the analyte to be detected can be coupled to, integrated into, or absorbed to the multifunctional copolymers. These multifunctional copolymer coatings are compatible with a variety of different established methods to detect, sense and quantify the target molecule in an analyte.

Abstract: The present invention relates to particulate material having a density of at least 2.5 g/ml, where the particles of the particulate material have an average diameter of 5-75 &mgr;m, and the particles of the particulate material are essentially constructed of a polymeric base matrix, e.g. a polysaccharide such as agarose, and a non-porous core material, e.g. steel and titanium, said core material having a density of at least 3.0 g/ml, said polymeric base matrix including pendant groups which are positively charged at pH 4.0 or which are affinity ligands for a bio-molecule. Possible pendant groups include polyethyleneimine (PEI), diethylaminoethyl (DEAE) and quaternary aminoethyl (QAE). The materials are useful in expanded bed or fluidized bed chromatography processes, in particular for purification of bio-macromolecules such as plasmid DNA, chromosomal DNA, RNA, viral DNA, bacteria and viruses.

Abstract: A compound possessing physiological activity is coupled to a styrene-glycidyl methacrylate polymer through a spacer. Compounds that may be used include receptors such as proteins, and 3-[(5-(2,3-dimethoxy-6-methyl-benzoquinonyl)]-2-nonyl-2-propionic a preferred spacer is an ethylene glycol diglycidyl ether derivative. Preferably, the whole surface of the styrene-glycidyl methacrylate polymer in microsphere form is covered with glycidyl methacrylate. The microsphere may be used for isolating and detecting substances such as proteins that bind to the coupled compound.

Abstract: The invention concerns a method for the purification of a target substance from a biological sample by immobilizing the target substance on a solid phase by means of a high affinity binding pair and subsequently eluting it by adding a partner of the binding pair in a free form. In addition reagent kits for carrying out the method are disclosed.

Abstract: Processes for preparing oligosaccharides, polysaccharides, glycolipids, glycoproteins, and other saccharide compositions are provided which involve the enzyme facilitated transfer of a preselected saccharide unit from a donor moiety to an acceptor moiety. In accordance with a preferred embodiment, saccharide compositions having a plurality of saccharide units are prepared by appending the saccharide units in iterative fashion to acceptor moieties which are themselves saccharide compositions prepared in accordance with this invention.

Abstract: The utility of soybeans having a composition of greater than 40% of the protein as beta-conglycinin and less than 10% of the protein as glycinin for making highly functional high beta-conglycinin compositions was discovered. The discovered ingredients are useful for mimicking the texturizing properties of casein while also maintaining or improving physiological benefits of soy protein ingredients (e.g., cholesterol and triglyceride lowering properties). The high stability of the high beta-conglycinin compositions against protein—protein aggregation reactions is valuable for creating good tasting beverages and beverage mixes. Cheese with good spreadability, gloss and smoothness was made using an enzyme-modified version of the new ingredient composition. Cheese with good firmness and meltability was also created using a different enzyme-treatment. High beta-conglycinin compositions were found to demonstrate excellent emulsifying and gelling properties in the pH region (5.5-6.

Abstract: A microsphere is prepared containing a compound possessing physiological activity coupled to a styrene-glycidyl methacrylate polymer through a spacer. Compounds that may be used include receptors such as proteins, and 3-[(5-(2,3-dimethoxy-6-methyl-benzoquinonyl)]-2-nonyl-2-propionic acid. A preferred spacer is an ethylene glycol diglycidyl ether derivative. Preferably, the whole surface of the styrene-glycidyl methacrylate polymer is covered with glycidyl methacrylate. The microsphere may be used for isolating and detecting substances such as proteins that bind to the coupled compound.

Abstract: Biological materials in a mixture of substances are separated, detected or quantified using magnetic spherically shaped cross-linked polyvinyl alcohol (PVAL) polymer particles ranging in size from 1 to 10 &mgr;m. The particles containing a coupled ligand are used to bind a biological material in a mixture of substances, and the particles containing bound biological material are isolated from the mixture. The particles are prepared by dispersing a magnetic colloid containing a magnetic material such as a ferromagnetic or superparamagnetic substance in an aqueous solution of polyvinyl alcohol containing reactive hydroxyl groups, adding the resultant mixture to an organic phase containing a mixture of at least two emulsifiers, and adding a water-soluble cross-linking agent such as a dialdehyde that reacts with the hydroxyl groups of polyvinyl alcohol to form the polymer particles.

Abstract: The present invention relates to the identification and purification of a herpes protease and a nucleic acid segment coding for two proteins. The first protein is the herpes protease which is able to cleave itself and also cleave the second protein. This protease is required for the assembly of the herpes virus capsid, therefore is essential for replication. The second protein has previously been designated as the family of proteins in viral infected cells, ICP35. The protease and its substrates are encoded by overlapping nucleic acid segments. This invention also relates to a promoter sequence for the second protein. Methods are presented of producing a viral protease, screening a protease inhibitor which may be used in a drug designed for the treatment of herpes disease, methods for treating herpes and other viral infections wherein the virus employs a protease substantially similar to the herpes protease, for capsid production.

Abstract: A method for the purification of human platelet heparanase using chromatographic techniques is described. The method comprises detergent-aided solubilization of a heparanase-containing fraction followed by purification of the heparanase therefrom by concanavalin A, Zn2+-chelating Sepharose, a Blue A or Reactive Red matrix, octyl agarose and gel filtration chromatography. Heparanase so purified has a molecular mass (Mr) of about 50 kDa and degrades both heparin and heparan sulfates.

Abstract: Magnetic particles are prepared and used for selective separation of affinity bound partners in solution. The magnetic particles have large magnetic moments which can be made such that different strengths of magnetic moments and/or different magnetic field dependencies can be used to allow for separation of several affinity partners simultaneously. Magnetic particles having different magnetic moments and different attached acceptor molecules move at different rates in a magnetic field. The magnetic particles have a first ferromagnetic layer having a moment oriented in a first direction, a second ferromagnetic layer having a moment oriented in a second direction generally antiparallel to the first direction and a nonmagnetic spacer layer located between and in contact with the first and second ferromagnetic layers.

Abstract: A method for preparing saccharide compositions which is reiterative and includes the following three steps. (i) A glycosyltransferase capable of transferring a preselected saccharide unit to an acceptor moiety is isolated by contacting the acceptor moiety with a mixture suspected of containing the glycosyltransferase under conditions effective to bind the acceptor moiety and the glycosyltransferase and thereby isolate the glycosyltransferase. The acceptor moiety is a protein, a glycoprotein, a lipid, a glycolipid, or a carbohydrate. (ii) The isolated glycosyltransferase is then used to catalyze the bond between the acceptor moiety and the preselected saccharide unit. (iii) Steps (i) and (ii) are repeated a plurality of times with the intermediate product obtained in the first iteration of the method being used as the acceptor moiety of the second iteration.

Abstract: The present invention relates to a method of screening for drug binding to serum proteins by: preparing at least two solutions each including a concentration of a serum protein and a concentration of a candidate drug, wherein the concentration of the candidate drug is different for each of the at least two solutions; exposing each of the at least two solutions to a light source; measuring fluorescent emission by the serum protein or a serum protein-candidate drug complex for each of the at least two solutions upon said exposing; and determining whether a change in fluorescence emission is measured for an increased concentration of the candidate drug, wherein the change in fluorescence emission indicates binding of the candidate drug to the serum protein. A kit useful for performing a fluorimetric screening of drug binding to serum proteins is also disclosed.

Abstract: The present invention includes the characterization of the major salivary protein or enzyme of the corn earworm Helicoverpa zea for triggering resistance to bacterial blight and frogeye leafspot in soybeans and for triggering resistance to insects in tomatoes. The invention includes an enzyme or a novel protein secreted from the salivary glands of certain insects including the saliva of species belonging to the order Hymenoptera and Lepidoptera. The regurgitant of Helocoverpa zea obtained from the functional salivary glands contains a protein that possesses glucose oxidase activity. The amino acid sequence of the protein is unique and when the protein is applied to plants, it triggers disease and insect resistance systematically. The physical and kinetic attributes of the enzyme are a pH of 7.0, a pI of 4.4 and a molecular weight of 88 kd. The km and Vmax of the enzyme for glucose is 26.9 mmol and 26.7 &mgr;mol min−1 mg−1, respectively.

Abstract: Activated support materials are provided containing oxirane or azlactone groups as substituents in linear polymers as activated groups. A base support containing hydroxyl groups is suspended in a solution containing cerium (IV) ions and a monomer containing an oxirane or azlactone group, and grafting polymerization is carrier out to produce a polymer containing oxirane or azlactone groups covalently bonded to the base support. Azlactone groups can be bonded to the base support via a thioether bond by using a base support containing thiol groups. The activated support materials can be used to prepare affinity supports containing an affinity ligand that is thiophilic or possesses a metal chelating group, or to prepare immobilized enzymes. The ligand can be iminodiacetic acid, or can be obtained by reacting an oxirane group of the support material with NaHS, and reacting the resultant product with divinylsulfone followed by reacting with mercaptoethanol.

Abstract: The use of calcium salts, magnesium salts or combination thereof in immunochemical methods for the determination of an analyte in a sample. These salts increase the specificity of such determinations, in particular, the background signal being reduced by addition of these salts in solid phase immunochemical methods.

Abstract: The present invention is to the discovery of a novel CD23 processing enzyme which is of importance in the human immune response and regulation of IgE production, and is a protein expressed on the surface of a variety of cells.

Abstract: A process for purifying thrombin-like proteases from snake venoms is described, which consists in freeing the proteases from impurities in three chromatographic steps: a) affinity or anion exchange, b) adsorption onto a glass matrix at alkaline pH values, and c) size exclusion gel or glass matrix at acidic pH values.

Abstract: A compound having a polysaccharide binding domain such as contained by a cellulose and essentially lacking in polysaccharidase activity is purified from other ingredients in a mixture using an affinity partition system. A mixture containing the compound is contacted with a system containing as a first phase an aqueous solution of oligosaccharide polymer such as cellulose and as a second phase a solution of a polymer such as a poly(ethylene glycol)-poly(propylene glycol) copolymer. The compound petitions into the first phase and binds to the oligosaccharide polymer, preferably with a Ka of 103 to 107, to form a complex. The complex is collected, and the compound is dissociated from the oligosaccharide polymer. The compound may be formed of a non-peptide chemical moiety or a peptide moiety linked to a polypeptide having the polysaccharide binding domain.

Abstract: A purified enzyme-I is obtained that participates in C-terminal amidation by acting on a peptide C-terminal glycine adduct to form a peptide C-terminal .alpha.-hydroxyglycine adduct. The enzyme has an optimum pH of about 5 to 7, an optimum temperature of 25 to 40.degree. C. and a molecular weight of about 25 kDa or about 36 kDa, and metal ions and ascorbic acid act as a cofactor. A purified enzyme-II is obtained that participates in C-terminal amidation by acting on the peptide C-terminal .alpha.-hydroxyglycine adduct to produce a C-terminal amidated compound. The enzyme has an optimum pH of about 5 to 6, an optimum temperature of 15 to 35.degree. C. and a molecular weight of about 40 kDa or about 43 kDa. Enzyme-I does not act on the peptide C-terminal .alpha.-hydroxyglycine adduct and enzyme-II does not act on the peptide C-terminal glycine adduct. The enzymes may be purified from a biological material such as horse serum by affinity chromatography using a peptide C-terminal glycine adduct as a ligand.

Abstract: An affinity chromatographic matrix for purification of a biological material is provided having an ionically charged polymeric ligand such as glycoaminoglycan non-covalently bound directly by an ionic bond to an oppositely ionically charged group on a chromatographic matrix. Having the ligand non-covalently bound to the matrix by an ionic bond, allows the ligand to be easily washed off the matrix and replaced for subsequent purifications without having to replace the matrix. A biological material in a crude mixture is purified by non-covalently binding the material to the bound ligand and dissociating the material from the ligand. Matrices that may be used include crosslinked agarose, crosslinked dextran, crosslinked cellulose, crosslinked dextran and bisacrylamide, or matrices based on silica or plastic polymers. The charged group may be a quaternary amine or diethylaminoethyl group.

Abstract: A two-phase partition system is provided for affinity separation of a composition containing a polysaccharide binding peptide from a mixture such as a fermentation broth. The peptide may be from an enzyme and lacking in polysaccharidase activity such as the binding domain of cellulase that binds to cellulose. The system contains a phase-forming oligosaccharide polymer such as a cellulose derivative to which the peptide binds with a Ka of 10.sup.3 M to 10.sup.7 M, and a phase inducing agent such as a polyethylene glycol polymer, or a salt present at sufficiently high concentration to induce phase separation. If the oligosaccharide polymer is thermoseparating, phase separation can be induced by heating. Using the system involves contacting a composition containing the peptide such as a fusion protein with the system, partitioning the composition into a phase containing the oligosaccharide polymer by binding to the polymer and recovering the polymer containing the bound composition.

Abstract: In a vertical down-flow fluid bed reactor, suspended particles in liquid proximal to an inlet in an uppermost part of the reactor are agitated to form a downward extending turbulent zone having vigorously moving particles and a non-turbulent zone distal to the inlet having essentially stationary particles in liquid below and adjoining the turbulent zone. In a vertical up-flow fluid bed reactor, an upward extending turbulent zone is formed proximal to an inlet in a lowermost part of the reactor and the non-turbulent zone is above the turbulent zone. The downward or upward extend of the turbulent zone is determined by the degree of agitation. The particles may contain an active substance and be in the form of a conglomerate of base particles having a desired density to control floatation or sedimentation. Particles in the turbulent and non-turbulent zones may be different such as having different specific gravities.

Abstract: A substance possessing physiological activity is coupled to a styrene-glycidyl methacrylate polymer through a spacer, and is used to isolate from a mixture a substance that can adhere to the substance possessing physiological activity. Preferably the polymer is in the form of a microsphere, the substance possessing physiological activity is 3-[(5-(2,3-dimethoxy-6-methyl-1,4-benzoquinonyl)]-2-nonyl-2-propionic acid or derivative thereof, and the spacer is an ethylene glycol diglycidyl ether derivative. The mixture may be a cell extract and the substance isolated a protein that is a receptor to the substance possessing physiological activity.

Abstract: Process for purifying serine proteases from a protein mixture by binding the serine protease to an immobilized polypeptide with the activity of an inhibitor DE-3 from Erythrina caffra, removing unbound components from the protein mixture, detaching the serine protease from the inhibitor and separating the immobilized inhibitor from the soluble serine protease and isolating serine protease which is characterized in that a polypeptide is used as the polypeptide which is the product of a prokaryotic or eukaryotic expression of an exogenous nucleic acid. This inhibitor is distinguished by an improved specific activity and is particularly suitable for the purification of plasminogen activators such as tissue plasminogen activators (t-PA and derivatives).

Abstract: Methods are disclosed for determining an analyte in a medium suspected of containing the analyte. One method comprises providing (1) combining a medium suspected of containing the analyte and a novel chemiluminescent compound, (2) combining a means for chemically activating the chemiluminescent compound; and (3) detecting the amount of luminescence generated by the chemiluminescent compound. The amount of luminescence generated is related to the amount of analyte in the medium. The chemiluminescent compound can be chemically activated by hydrogen peroxide. Compositions and kits are also disclosed.

Abstract: A reiterative method for obtaining the glycosyltransferases useful for glycosyltransferase-catalyzed synthesis of a saccharide composition by serially bonding preselected saccharide units onto an immobilized acceptor moiety which is one member selected from the group consisting of proteins, glycoproteins, lipids, glycolipids and carbohydrates.

Abstract: The present invention provides the identification and characterization of two components of a recombinant preparation of DNase. These components are the purified deamidated and non-deamidated human DNases. Taught herein are the separation of these components and the use of the non-deamidated species as a pharmaceutical per se, and in particular in compositions wherein the species is disclosed within a plastic vial, for use in administering to patients suffering from pulmonary distress.

Abstract: A method is provided for preparing a low density porous rigid fused-fiber matrix having a density of between about 3.5 and 5.5 pounds/cubic foot and a free volume of between about 90-98 volume percent for use as a cell-culture substrate or implant material, or in chromatographic separation of blood cells. The method is carried out by forming a slurry containing (I) silica, alumina or silica and alumina fibers having thicknesses between about 0.5 and 20 .mu.m and lengths between about 1 and 10 mm, and having a fiber:liquid weight ratio of between about 1:25 to 1:70, (ii) a thickening agent to give the slurry a viscosity between about 1,000 and 25,000 centipoise, (iii) boron nitride particles between about 2-12 percent by weight of the total fiber weight, and (iv) a dispersing agent when the slurry contains silica fibers, allowing the slurry to settle in a mold to produce a fiber block, drying the fiber block, and heating the dried fiber block to at least about 2200.degree. F.

Abstract: Perfluorocarbon polymer-based matrices are coated with a hydrophilic polymer for use in bioaffinity separations. Coating is carried out by dispersing porous particles of inert perfluorocarbon polymer in a water-miscible organic solvent such as acetone or tetrafydrofuran to wet surfaces of the particles, forming a dispersion of the wetted particles in an aqueous solution of hydrophilic polymer such as poly(vinyl alcohol) containing a plurality of hydroxyl groups, at least one being at an end of a polymer chain, to adsorb the hydrophilic polymer onto the wetted surfaces of the particles, admixing a homobifunctional cross-linking agent such as glutaraldehyde with the particles to cross-link the hydrophilic polymer, activating hydroxyl groups on the surface of the cross-linked hydrophilic polymer and covalently bonding a ligand or ligand binder to the activated hydroxyl groups.

Abstract: An affinity support is provided containing a high flux semipermeable hydrogel membrane surface-modified with an affinity ligand, such as a protein which may be an antibody or enzyme, or a cell receptor complement, useful for affinity separation of biological macromolecules, including insoluble proteins, cells, and cell fragments, from solution. The exclusion limit (molecular weight cut-off) of the matrix is selected to substantially restrict immobilized protein or other ligand to the surface thereof for maximization of available ligand binding capacity. The exclusion limit is also selected to permit reagent(s) used for the matrix/ligand linkage to penetrate into and form covalent bonds with the membrane on the interior surfaces of the membrane for optimizing packing densities of the affinity ligand exterior to the membrane.

Abstract: Device for the treatment of nucleic acids from the sample, comprising a first reaction chamber for separating the nucleic acids from other sample components, and a second reaction chamber for the amplification of the nucleic acids, connected to said first reaction chamber via a controllable transport path.

Abstract: A method is provided for preparing an asparaginyl endoproteinase from the seeds of soybean, ginkgo and rice which have been collected between an early growing stage and ripening. The method comprises the steps of dialyzing an extract of the seeds against an acidic buffer of pH 4.0 to 6.0, ammonium sulfate precipitation, hydrophobic chromatography and gel filtration. The resulting asparaginyl endoproteinase cleaves glycinin between the C-terminal amino acid residue of the acidic subunit region, Asn, and the N-terminal amino acid residue of the basic subunit region, Gly or Asn.

Abstract: A gene is provided encoding a DNA sequence for the expression of parathion hydrolase and a methionine analog thereof. In a preferred embodiment, a parathion hydrolase gene encoding sequence is obtained in which twenty-eight amino acids are deleted from the N-terminal amino acid sequence of parathion hydrolase. Parathion hydrolase or its analog is produced by transforming a host microorganism such as Escherichia, Bacillus or Streptomyces with the gene, culturing the microorganism in a culture medium and purifying parathion hydrolase therefrom. Highly purified soluble parathion hydrolase is produced by a purification method without using detergents such as Triton X-100 and Tween 20. Parathion hydrolase of enhanced activity is produced by adding cobalt or zinc or a mixture thereof to the culture medium in which the host microorganism is cultured.

Abstract: This invention relates to an apparatus containing specific binary combinations of glycosyltransferases, for the synthesis of specific saccharide compositions such as, for example, oligosaccharides, polysaccharides, glycolipids, and glycopeptides.

Abstract: Echinocandin B deacylase is purified to near homogeneity from Actinoplanes utahensis by a process comprising, in order, extracting the soluble enzyme, heating to 60.degree. C., hydrophobic interaction chromatography, (NH.sub.4).sub.2 SO.sub.4 fractionation, gel filtration, cation exchange chromatography, dye-ligand chromatography, gel filtration, and cation-exchange chromatography.

Abstract: A dipeptidylaminopeptidase (dDAP enzyme) and a method for isolating it from the culture medium of the cellular slime mold, Dictyostelium discoideum have been presented. The isolated dDAP enzyme has a pH optimum of about 3.5 and a mass of about 225 kDA. The dDAP enzyme has an activity which is somewhat similar to both DAP-I and DAP-III from this organism. Methods for using the dDAP enzyme to remove dipeptides from the N-terminus of recombinantly produced precursor proteins or peptides are also presented.

Abstract: Aspergillus niger acid protease is prepared by treating whole glucoamylase product containing a mixture of acid protease and glucoamylase with an anion exchange medium which selectively removes acid protease from the glucoamylase, producing a glucoamylase product which is protease-free. The acid protease is then recovered from the anion exchange medium by elution with high salt or low pH solution.

Abstract: A .gamma.-phosphate-linked ATP affinity column for the purification of a protein kinase is provided. In addition, a method for purifying protein kinases using the .gamma.-phosphate-linked ATP affinity column comprising the steps of 1) linking an ATP linking moiety to an affinity column via the .gamma.-phosphate position of ATP; 2) applying a sample of protein kinase to the .gamma.-phosphate linked ATP solid support so as to bind the protein kinase to the .gamma.-phosphate-linked ATP; 3) washing the bound support with a salt which will not affect the binding of the bound protein kinase; and 4) eluting the bound protein kinase with an ATP salt so as to obtain homogenous protein kinase is provided.

Abstract: An ultra-pure, clear thrombin solution having a high specific activity is described as well as a method of manufacture.

Abstract: A chromatographic process for the copurification of chondroitinase proteins useful in ocular surgery for non-surgical disruption of chondroitin sulfate, the molecule which mediates the attachment between the retina and vitreous body in the human eye. The process involves the use of ion exchange resins in conjunction with an affinity elution with chondroitin sulfate to afford copurification of the proteins.

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.

Abstract: A Coffea canephora-D-galactosidase isozyme is purified by extracting a supernatant from Coffea beans containing the isozyme, extracting tannin from the supernatant, and isolating the isozyme. Preferrably, the supernatant is extracted by exposing the supernatant to insoluble polyvinylpolypyrrolidone in an amount sufficient to remove the tannin from the supernatant by forming hydrogen bonds with the tannin.