Abstract: Method for the bioluminescent dosing of enzymes, substrates or enzymatic inhibitors, characterized in that there is used a transparent support made of synthetic plastic material previously treated with a charged hydrophilic substance for the Fixation by adsorption of an enzymatic system containing at least a luciferase. The invention extends to a support of the polystyrene type treated by a charged hydrophilic substance on which an enzymatic system containing at least a luciferase is fixed by adsorption.

Abstract: Enzymatic catalysts are produced from mineral particles coated with chitosan. The enzymes may be directly absorbed onto the surface after a cross-linking treatment to stablize the coating or immobilized by chemical bonds after a treatment with a bifunctional reagent, for example with glutaraldehyde. The stability of the biocatalyst is advantageously increased by retreatment with a bifunctional reagent after immobilization of the enzyme. Enzymes sensitive to glutaraldehyde are fixed by a carbodiimide. The catalysts thus prepared are insoluble in aqueous solutions. The particles are rigid and incompressible and are suitable for use in fixed-bed and fluidized-bed reactors. The immobilization yield may reach 90% depending on the circumstances.

Abstract: A water-soluble peroxidase derivative obtainable by the oxidation of peroxidase with periodic acid or with a salt thereof and binding to a water-soluble polymer selected from polysaccharides, polyethylene glycols, polyvinylpyrrolidones and polyacid anhydrides. Also provided is a process for the preparation of this water-soluble peroxidase derivative and for the use of this water-soluble peroxidase derivative for the determination of hydrogen peroxide.

Abstract: Modified polymers based on polyvinylene carbonate and/or polyhydroxymethylene are prepared containing covalently bonded units derived from particular alkoxylated compounds. Polymerization is carried out in the presence of the alkoxylated compounds and a particular dispersion stabilizer. The modified polymer is suitable as a carrier for biologically active substances or for affinity chromatography.

Abstract: A method for enzyme immunoassay includes contacting under binding conditions a liquid suspected of containing an analyte, an antianalyte affixed to a solid support and a tracer having an enzyme conjugated thereto. A bound fraction is separated from the liquid and incubated in a second liquid with a masked ligand. The masked ligand is converted by the enzyme on the bound fraction to give free lignad which binds to an antiligand. A signal system, such as a signal enzyme and substrate therefor, or a label-loaded vesicle and vesicle lysing agent, is added to generate a signal used to detect or measure the analyte in the liquid. The invention includes a kit of materials useful in performing the assay of the invention.

Abstract: A water-insoluble, crosslinked polyacrylamide copolymer is prepared having recurring monomeric residues of acrylamide, bisacrylamide or analogs thereof, and N,N'-bisacrylylcystamine or analogs thereof. The crosslinked copolymer can be formed into discrete particles having crosslinking disulfide groups on their external surface which can be reduced to provide activated particles having exposed chemically active sulfhydryl groups. The activated polyacrylamide particles are free of residual amino and carboxylic acid groups and can be reacted with suitable haptens and other biologically active substances to provide insolubilized forms thereof.

Abstract: An immunogenicity increasing carrier substance for immunogenic determinants is composed of particles derived from gram-negative cells substantially devoid or depleted of their natural immunogenic determinants, more particularly prepared by stripping the O-antigen and core region off the lipid A by chemical cleavage (acid hydrolysis). These socalled "naked bacteria" are chemically treated with a linking reagent to provide covalently bonded intermediate linking moieties while preserving the adjuvant effect of the particles. The linking moieties have functional groups. These serve for the covalent bonding of selected haptens or antigens comprising the desired immunogenic determinants against which antibodies are to be elicited in living antibody-producing cells.

Abstract: A bridging molecule carrying a drug, or a label such as a fluorophore, which adds across disulfide bonds of molecules, particularly proteins, and methods of manfacturing and using the bridging molecules, are disclosed. The bridging molecule is reactive with sulfhydryl groups formed by the reduction of disulfide bonds of the protein. The functional groups of the bridging molecules are typically --SH groups.

Abstract: A porous chemically activated medium having a low affinity for peptide group-containing materials is provided comprising a porous polymeric medium having a low affinity for peptide group-containing materials covalently bound to a residue of an activating agent which is capable of reacting with an acceptor molecule.

Abstract: Thiol gels are prepared from hydroxyl containing polymers by forming a 2-fluoro-1-methylpyridoxy derivative of the polymer followed by reacting the derivative with sodium dimethyldithiocarbamate and reducing to produce a sulfhydryl substituted polymer. Alternatively, the derivative is reacted with dithiothreitol to produce a sulfhydryl substituted polymer having free sulfhydryl groups spaced from the polymer. The thiol gel can be activated by means of 2,2'-dipyridyl disulfide and reacted with a free sulfhydryl containing biologically active ligand in order to provide an insolubilized ligand which can be used for various purposes including use as an immobilized biologically active material or use as a matrix in covalent chromatography.

Abstract: A bioaffinity separation method is provided along with a solid affinity support utilized in that method. Additionally, immobilized enzyme systems are provided for use as enzyme electrode systems. The support is based on an inert perfluorocarbon polymer carrier with ligands or binders attached to its surface. The ligand, binder or enzyme is preferably modified by attaching a perfluorocarbon anchor group, and the modified ligand, binder or enzyme is attached to the carrier through the anchor group. Methods for preparing such supports and their use in capturing target molecules from samples and in analytical applications are also provided.

Abstract: There are provided magnetic and non-magnetic agarose and agar polyaldehyde beads with diameters ranging from 40 microns up to 1 cm, and processes for the synthesis of such beads. The polyaldehyde compounds e.g. polyacrolein, polymethacrolein or polyglutaraldehyde, were used as microspheres or as powders. The agarose-polyaldehyde beads are capable of covalently binding in a single step, through their aldehyde groups, compounds containing primary amino groups or thiol groups, such as proteins, antibodies, enzymes and drugs. The beads are useful for various biological applications e.g. affinity chromatography, hemoperfusion, ion exchange resins, cell labeling, diagnostic purposes and cell separation.

Abstract: The present invention relates to a continuous process for the enzymatic preparation of isomaltulose. A periplasmatic sucrose-mutase is produced by fermentation of microorganisms which form sucrose-mutase. The cell-free crude enzyme extract is prepared by digestion of the cells and by cross-flow microfiltration. In a single-stage, simultaneous purification and immobilization of the sucrose-mutase from the cell-free crude extract conditioned by diafiltration is obtained by selective bonding to an anionizable carrier matrix. Direct conversion of sucrose into isomaltulose is produced by the sucrose-mutase bonded to the anionizable carrier matrix, preferably in cartridge or cartouche form.

Abstract: A composite article is prepared having in sequence a fibrous support which has been subjected to a surface treatment to provide binding sites thereon, a layer of a protein immobilizer compound, and a biologically active protein such as enzyme. The surface treatment can be carried out by coating the surface with an inorganic oxide or by subjecting the surface to plasma treatment.

Abstract: Materials and a method of their preparation are described, for use in immobilizing soluble, biologically active compounds. The material for immobilizing includes a hydrophobic, water insoluble substrate having a coating of a mixture of different amphiphilic lipoid compounds thereon. The lipoid compounds are oriented so that the hydrophilic portions are distal to the substrate and free to bond with water soluble proteins and carbohydrates.

Abstract: A microorganism or enzyme is mixed with an alginate and a silica sol in the presence of water to obtain a liquid mixture having a pH of 3 to 10 and containing an alginate concentration of 0.5 to 3.5% (w/v) and a silica concentration originating from the silica sol of 0.5 to 35% (w/v). The mixture is contacted with a gelling agent in the form of an aqueous solution to obtain a gel containing the microorganism or enzyme.

Abstract: A magnetic-polymer particle, useful in immunoassay techniques and various other biological/medical applications is produced by coprecipitation of transition metals in the presence of a polymer having available coordination sites. These particles are capable of forming stable aqueous suspensions and may be easily resuspended after agglomeration.

Abstract: Glucoamylase is immobilized with a carbodiimide on a polymer having amino or carboxy functional groups. When the polymer contains amino groups, the glucoamylase is treated with the carbodiimide and then mixed with the polymer or the carbodiimide is added to a mixture of the glucoamylase and the polymer. When the polymer contains carboxy groups, the polymer is treated with the carbodimide and then mixed with the glucomylase. The polymer is produced from acrylic acid, metharylic acid, acrylamide, methacrylamide, N-amino-ethyl-acrylamide and/or N-amino ethyl methacrylamide and a cross-linking agent of the acrylic or allylic type.

Abstract: A carrier for the cultivation of animal and/or human cells in a fermenter, comprises a polymer base body of a solid formed into a pressure-resistant matrix, with a capillary system for carrying the liquid cell culture medium therethrough, with no cell growth occurring therein, and with interstices for three-dimensional cell growth therein, growth factors for the cells being covalently bonded to the boundary surfaces of the interstices. The carrier can provide a high level of cell density of the order of magnitude of 10.sup.9 cells/ml, by three-dimensional cell growth.

Abstract: Enzymes are fixed on carriers with a bis-dithioester capable of combining with the enzyme and with the carrier. The bis-dithioester has at each of its ends a group capable of reacting with an aimine function. The enzyme may be contacted with the bis-dithioester before the enzyme and carrier are combined. The carrier is an inorganic or organic solid.

Abstract: A hydrophilic coating photopolymerizable in air, capable of fixing bioactive molecules is formed from a mixture of acrylic or methacrylic monomers containing at least 20% by weight of an approximately quimolecular mixture of acrylic acid and an N-dialkylated amino-alcohol acrylate or methacrylate.

Abstract: Heterobifunctional reagents that are cleavable under mildly acidic conditions are disclosed. The reagents include an acid cleavable cyclohexene-1,2,-dicarboxylic acid monoamide group. Also disclosed are methods of making the cross-linkers, as well as methods of using the cross-linkers, e.g., to deliver a biologically active substance across the membranes of selected cells in a heterogeneous cell population; once inside the cell the active substance is released, intact, by the transient, mild acidity of certain cell structures. Finally, a method of characterizing complex multi-chain protein structures is disclosed.

Abstract: Enzyme-containing microbial cells are contacted with a macro-porous non-ionic resin under reduced pressure below atmospheric to cause the cells to become incorporated into pores of the resin. The resin is then contacted with a multi-functional, amine reactive material to immobilize the cells within the pores. Prior to contacting with the resin, cells may be disrupted to produce a system containing cell fragments, intact cells and solubilized enzyme. A preferred enzyme is glucose isomerase.

Abstract: A matrix useful for immobilizing organic ligands, such as biologically active materials, is prepared by reacting a hydroxyl group containing polymer with polyethyleneimine.

Abstract: A bridging molecule carrying a drug, or a label such as a fluorophore, which adds across disulfide bonds of molecules, particularly proteins, and methods of manufacturing and using the bridging molecules, are disclosed. The bridging molecule is reactive with sulfhydryl groups formed by the reduction of disulfide bonds of the protein. The functional groups of the bridging molecules are typically --SH groups.

Abstract: A derivative of a fibrinolytic enzyme in which the catalytic site on the enzyme which is responsible for fibrinolytic activity is blocked by a human protein attached thereto by way of a reversible linking group.

Abstract: A reaginic agglutination test for syphilis-associated antibodies is disclosed. The test uses an antigen reagent that comprises a buffered aqueous suspension of cardiolipin antigen ionically coupled to latex particles via a polypeptide bridge. Positive sera react with the antigen reagent and yield an agglutination pattern characterized by medium to large aggregates. Negative sera yield no agglutinated particles.

Abstract: The process for the separation of antiVIII:C antibodies present in a liquid consists of contacting said liquid with a solid support constituted by a polymer or a copolymer having in its chain substitutable groups, whereof at least part is substituted by groups having an affinity for antiVIII:C antibodies and a selectivity for antiVIII:C antibodies compared with other immunoglobulins and then separating the liquid from the support on which have been adsorbed the antiVIII:C antibodies.Preferably use is made of polystyrene, to which have been fixed groups --SO.sub.3 Na and --SO.sub.2 Glu, --SO.sub.2 Threo, --SO.sub.2 OHPro or SO.sub.2 Lys. These supports can be used for ex-vivo purification in the column of the blood plasma of a type A hemophiliac.

Abstract: A derivative of a fibrinolytic enzyme in which the catalytic site on the enzyme which is responsible for fibrinolytic activity is blocked by a human protein attached thereto by way of a reversible linking group.

Abstract: A modified silica material which comprises silica covalently bonded to a synthetic polymer, the synthetic polymer being made from (a) a polymerizable compound capable of being covalently coupled directly to said silica, and (b) one or more polymerizable compounds containing (i) an ionizable chemical group, (ii) a chemical group capable of transformation to an ionizable chemical group, (iii) a chemical group capable of causing the covalent coupling of the compound (b) to an affinity ligand or a biologically active molecule or (iv) a hydrophobic compound.

Abstract: Adsorbents, which are suitable for the affinity chromatography of proteins, especially of interferons, and which consist of an electroneutral carrier matrix, --O--CH.sub.2 --CH(OH)--CH.sub.2 --NH- groups as the spacer and a triazine coloring substance bonded to the spacer.

Abstract: An improved process of performing analysis methods based upon biospecific affinity reactions in heterogeneous systems, wherein a ligand dissolved in a liquid is reacted with a receptor immobilized on a carrier, characterized by utilizing as the carrier a porous matrix in the form of a material body having an open capillary pore system capable of absorbing and retaining liquid phase and having such limited void dimensions that the total reaction rate of the biospecific reaction or reactions within the pore system between ligand and immobilized receptor will be at least substantially independent of the diffusion of the ligand in the liquid phase. A porous matrix for use in said process has the characteristics mentioned above.

Abstract: A membrane is produced by dissolving, in a water-miscible solvent, a water insoluble copolymer of acrylontrile with at least one monomer selected from the group consisting of an aminostyrene, a vinyl pyridine and an N-hydroxy-containing-substituent-acrylamide, casting said solution to form a thin layer of solution, contacting said solution with water thereby to coagulate the copolymer into a film, and washing away from the copolymer film the solution of solvent and water. The amine or hydroxy group of the copolymer is then activated and coupled with a ligand such as glucose isomerase, chymotrypsin or Protein A. The coupled membranes can then be used for biological separations and reactions.

Abstract: The adsorbent consists of a solid phase, completely or partially penetrated by or surface-coated with a hydrophilic molecular polymer netting to which has been bound substituents or cross bridges having chain sequences of the structureX--CH.sub.2 --CH.sub.2 --SO.sub.2 --CH.sub.2 --CH.sub.2 --S--Ywhere X is ether oxygen, a thioether sulfur atom or a nitrogen atom and Y is a substituted or unsubstituted alkyl, aryl or heteroaromatic group. The solid phase consists of particles with a diameter of less than 1 mm and the molecular polymer netting consisting of a cross-linked polyhydroxy polymer such as a polysaccharide, preferably a polygalactane such as agar or agarose or a cross-linked polyamide, e.g. polyacryl amide. Y can be hydroxy alkyl or mercapto alkyl, or phenyl alkyl or phenyl substituted with halogen or nitro groups. The adsorbent is prepared by first converting in a known manner a hydrophilic polymer containing OH and/or CONH.sub.

Abstract: A chemically active membrane having a large surface area is provided in which a hydrophilic, microporous, skinless, polyamide membrane is chemically bound to a residue of an activating agent which is capable of reacting with a biologically active material.The chemically active membrane, formed by reacting a hydrophilic, microporous, skinless, polyamide membrane with an activating agent may be used to prepare a biologically active membrane having a large surface area which comprises an acceptor molecule such as a monoclonal antibody, a polyclonal antibody, an antigenic substance, a glycoprotein, Protein A, a lectin, a carbohydrate, an enzyme substrate, a cofactor, an inhibitor, a hormone, an IgG class of immunoglobulin, a carrier protein, a receptor, heparin, a coagulation factor, or a histone covalently bound to the hydrophilic, microporous, skinless, polyamide membrane by reacting the chemically activate membrane with the acceptor molecule.

Abstract: A porous, bead-like copolymer which can be employed to excellent effect for the fixation of enzymes is obtained by suspension polymerization of monomeric oxiranylalkyl compounds and/or 2-aziridinylalkyl compounds, n-valent crosslinking agents, N-vinylamides, heterocyclic 5-membered ring compounds or 6-membered ring compounds, having a polymerizable olefinic group in each case, and polymerizable quaternary ammonium salts.

Abstract: A fibrinolytically active protein conjugate comprising at least one optionally blocked fibrinolytic enzyme linked by way of a site other than the catalytic site responsible for fibrinolytic activity to at least one human protein.Processes from making the conjugates and pharmaceutical compositions containing them are also described.

Abstract: The invention relates to a process for the immobilization of compounds comprising nucleophilic groups.According to the process of the invention a polymer comprising an amido group is swollen in a buffer. For this purpose any buffer is suitable which contains no amino, sulfhydryl and/or hydroxy groups in the molecule. To the swollen polymer a solution of a quinone - preferably p-benzoquinone - in a water miscible organic solvent is added and the mixture is activated at 273.degree.-343.degree. K. for 0.5-48 hours - preferably for 24 hours. The unreacted quinone is washed out with an aqueous solution of a water miscible organic solvent and thereafter water and/or a buffer. To the purified gel at a pH value between 3 and 11 - preferably between 6 and 8 - a buffered solution of a compound comprising a nucleophilic group is added, the suspension is incubated at 260.degree.-313.degree. K. for 24 hours, the gel is separated and the non-bound compound comprising a nucleophilic group is removed by washing.

Abstract: Bifunctional aromatic compounds are employed as rigid coupling compounds for coupling one organic compound to another. For example, paranitrophenylisocyanate may be employed as a coupling compound for coupling thyroxine to a fluorescent dye to form a tracer for use in an assay.

Abstract: A modified polysaccharide material which comprises: (1) polysaccharide covalently bonded to a synthetic polymer; (2) the synthetic polymer being made from (a) a polymerizable compound which is capable of being covalently coupled directly or indirectly to said polysaccharide, and (b) one or more polymerizable compounds containing (i) an ionizable chemcial group, (ii) a chemical group capable of transformation to an ionizable chemical group, (iii) a chemical group capable of causing the covalent coupling of the compound (b) to an affinity ligand or a biologically active molecule or (iv) a hydrophobic compound.

Abstract: Apparatus for immobilizing biological cells based on the dielectric properties of biological cells. An inhomogeneous electric field emanating from a grid point location or other contact area is created and attracts the biological cell into contact therewith. Appropriate controls over a series of grid points permits controlled inter-grid point movement of cells thereby permitting sequential testing or sorting processes to be carried out.

Abstract: Plastics surfaces are preactivated for immobilizing organo-chemical and biologic materials by coating the surfaces with a polypeptide which consists of a defined composition of hydrophobic amino acids and amino acids having side chains capable of activation and coupling. After activation of the plastics surfaces having been coated in this manner with various coupling reagents, it is possible to firmly fix organo-chemical and biologic materials. The materials having been immobilized in this manner are extremely stable to various reaction conditions.

Abstract: A method of chemical analysis is disclosed which employs a release tag compound of general formulaRx--Re--Sin which Rx is a reactivity group capable of forming a covalent bond with another molecule, Re is a release group capable of being cleaved, and S is a signal group. The release tag is covalently bonded to a substance of interest which is to be determined in the course of a chemical analysis for an analyte, release group Re is cleaved at an appropriate point in the analytical procedure, and signal group S is determined, thereby determining the substance of interest. Where the substance of interest is the analyte, determination of S determines the analyte. Where the substance of interest is not the analyte but is related to the analyte concentration, determination of S allows indirect determination of analyte.

Abstract: Carboxypeptidase B enzyme is isolated by a process wherein homogenized and autolyzed pancreas is subjected to a thermal treatment, and carboxypeptidase B is separated by fractionation with ammonium sulfate. This process produces carboxypeptidase B having satisfactory specific activity while eliminating the use of pancreas cell fluid or an acetonous powder from pancreas as a starting material. The isolated carboxypeptidase B may be immobilized on a polymer gel containing carboxy groups activated with a carbodiimide.

Abstract: Derivatives of a nonimmunogenic plasminogen activator which comprises at least one polyalkylene glycol group chemically bonded with at least one coupling agent to amino acid side chains of said plasminogen activator, wherein said polyalkylene glycol has a molecular weight of about 200-20,000 and is unsubstituted or is substituted with one or more alkyl, alkoxy or alkanoyl groups or a mixture thereof.The plasminogen activator derivatives have an extended circulating life in the mammalian bloodstream and also inhibit the formation of thrombus in the same.

Abstract: A process for removing immunosuppressive substances from the blood serum of cancer patients comprises contacting the serum at a temperature below room temperature and preferably not greater than 5.degree. C., with Protein A of S. aureus immobilized on a substrate. Conducting the immunoabsorption at low temperature reduces the amount of anaphylatoxins produced in the process and thereby reduces the side effects of immunoperfusion treatment. An apparatus for low-temperature immunoperfusion treatment comprises a chamber for containing the immunoabsorbent, inlet and outlet tubes connected to the chamber, and a cooling jacket surrounding the chamber and at least a portion of the inlet tube.

Abstract: A photo-cleavable compound for delivery and release of a biologically active substance to selected target cells; the compound includes a binding partner for a specific cell-surface receptor of those target cells, the biologically active substance to be delivered, and a photo-cleavable bridge between the binding partner and the biologically active substance. When the compound is exposed to a heterogeneous population of target and non-target cells, it binds selectively to the receptors on the surface of the target cells. Exposing the compound to light of selected wave length cleaves it, yielding the active substance.

Abstract: A solid-phase immunoassay support is provided by treating a test tube liner such as a preformed soft gelatin capsule half with a coupling solution comprising a fixative, a bifunctional coupling agent and a bioactive protein which selectively reacts with the substance which is the subject of the immunoassay. The support is particularly applicable to the radioimmunoassay (RIA) technique for determining the presence and concentration of minute amounts of protein antigens in biological fluid samples and can be readily inserted in an appropriately sized test-tube for easy and quick assays.

Abstract: Amino-sulfhydryl cross-linking reagents that are cleavable under mildly acidic conditions are disclosed. Also disclosed are methods of making the cross-linkers, as well as methods of using the cross-linkers, e.g., to deliver a biologically active substance across the membranes of selected cells in a heterogeneous cell population; once inside the cell the active substance is released, intact, by the transient, mild acidity of certain cell structure. Finally, a method of characterizing complex multi-chain protein structures is disclosed.

Abstract: A nylon matrix is partially hydrolyzed to form free amine end groups which are then linked to a protein molecule, e.g., an antigen or antibody, by a divalent radical which is the residue of a diimide molecule. The nylon carrying the protein can then be used in conventional manner in an immunoassay or an enzyme assay.