Abstract: The present invention provides a restriction endonuclease which recognizes palindromic sequences ##STR1## where C* is methylated, and cleaves these sequences at the position indicated by the arrows. This endonuclease is preferably from a microorganism of the genus Kluyvera. The present invention also provides a process for obtaining this new restriction endonuclease and a method for using the endonuclease.

Abstract: A reagent comprising a fraction obtained from plasma of an insect such as silkworm larvae and capable of reacting specifically with .beta.-1,3-glucan or peptidoglycan can be used for determining .beta.-1,3-glucan or peptidoglycan.

Abstract: A process for the isolation and separation of lysozyme and avidin from egg white is carried out by:step (a) contacting egg white with a weakly acidic cation exchange resin whereby lysozyme and avidin are adsorbed on to the resin; separating the resin from the egg white and washing the resin to remove residual egg white therefrom; and contacting the washed resin with a low ionic strength eluting buffer whereby lysozyme is eluted from the resin while avidin remains adsorbed on the resin; andstep (b) repeating the complete procedure defined in step (a) for two or more times; andstep (c) finally contacting the resin containing accumulated adsorbed avidin with a high ionic strength eluting buffer whereby avidin is eluted from the resin.Lysozyme and avidin are both commercial products useful, for example, in pharmaceutical applications.

Abstract: Methods for recovering t-PA from a liquid medium are disclosed. The methods comprise contacting a liquid medium with at least one substrate capable of effecting a separation of intact t-PA from degraded t-PA thereafter recovering the intact t-PA free from other unrelated protein. The present invention also provides compounds produced by this method, compounds comprising intact one-chain t-PA and pharmaceutical compositions containing them and methods for using such compositions.

Abstract: A novel plasminogen activator having specific properties. The plasminogen activator can be prepared by subjecting a human kidney or human blood vessel to an extraction treatment with an ammonium thiocyanate solution and then allowing the resultant extract to pass through a column of ion exchanger, a metal chelate column, a column of L-arginine or an arginine derivative supported on a carrier, or a column of a carrier having properties as a molecular sieve to purify the same. The plasminogen activator exhibits a strong thrombolytic activity and is useful as an active ingredient of a thrombolytic composition accompanied with a minimum of side effects.

Abstract: A process for the purification of alcohol oxidase from whole cells of Pichia pastoris grown on methanol by the sequential steps of autolysis, crossflow filtration, ultrafiltration and recrystallization.

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 through a highly fluorinated isocyanate anchor group. Methods for preparing such supports and their use in capturing target molecules from samples and in analytical applications are also provided.

Abstract: A member of a bioaffinity binding pair is immobilized on a plastic surface by coating the surface with poly(ethyleneimine) derivatized with a hydrophobic group, and covalently coupling the member to the coated surface. The immobilized member may be used in immunoassays or bioaffinity separations. The derivatized poly(ethyleneimine) is preferably tosyl poly(ethyleneimine).

Abstract: An isoenzyme from tomato, acid phosphatase-1 isoenzyme (Apase-1.sup.1), has ben purified to homogeneity and is subjected to amino acid sequencing and used to prepare anti-Apase-1 antibodies. The amino acid sequence permits design of probes to recover Apase-1.sup.1 -encoding cDNA; the antibodies are also useful for this purpose. The cDNA is useful to recover the genomic DNA encoding Apase-1.sup.1, which can then be used in walking or jumping techniques to recover the genomic DNA which confers nematode resistance, since this DNA resides immediately adjacent to the Apase-1.sup.1 gene on chromosome 6 of Lycopersicon esculentum.

Abstract: Disclosed is a method of recovering tissue plasminogen activator from media and cell extracts. The method comprises contacting the tPA-containing solution with a silicaceous matrix material comprising covalently bound polymer having plural anionic groups. Selective elution of the tPA can produce eluants of high specific activity. The method can succeed in recovering greater than 90% of the tPA activity from the crude solution.

Abstract: This invention relates to a process of preparation of nucleotide polymers wherein a lysate of a bacterial culture is passed successively through three columns (ion exchange resin, hydrophobic resin and molecular sieve), whereby there is obtained a substantially pure polynucleotidephosphorylase solution.Polymerization of this agent leads to non toxic and non pyrogen products.This invention relates also to products thus obtained and to therapeutic compositions containing the same.

Abstract: Disclosed is a method of recovering urokinase compounds from media and cell extracts. The method comprises contacting the urokinase compound-containing solution with a silicaceous matrix material comprising covalently bound polymer having plural anionic groups. Selective elution of the urokinase compound can produce elutants of high specific activity. The method can succeed in recovering greater than 90% of the urokinase activity from the crude solution.

Abstract: For selectively and reversibly adsorbing tPA, pro-tPA or mixtures thereof, from an aqueous medium, for example, derived from a melanoma cell culture, the aqueous medium is intimately contacted with an affinity reagent comprising an immobilized Kunitz-type inhibitor, substantially as occurs in and is extractable from seeds of an Erythrina species, e.g., Erythrina latissima, having Erythrina species, the following combination of characteristics:(a) it is an inhibitor of the Kunitz-type;(b) it inhibits trypsin;(c) it inhibits plasmin;(d) it has no effect on urokinase;(e) it inhibits tPA; and(f) in its immobilized form, it is a selected adsorbent for tPA and its precursor pro-tPA, to thereby cause the tPA, pro-tPA or mixtures thereof to become selectively adsorbed on the affinity reagent.

Abstract: A monoclonal antibody which is specific to human protein C which has calcium bound at the gammacarboxyglutamic acid(Gla) domain and does not recognize human protein C which is not bound to calcium at the Gla domain. The monoclonal antibody is used in an immunoassay method for determining human protein C having a Gla domain and in a method for recovering human protein C having a Gal domain.

Abstract: A mixture containing a single-chain tissue plasminogen activator (tPA) and/or double-chain tPA is brought into close contact with a column carrying an immobilized Erythrina trypsin inhibitor as an affinity agent. Adsorbed protiens are eluted with eluents having different pHs with or without arginine or benzamidine, so that single-chain tPA is obtained in the eluent with a pH at least 4.5 and double-chain tPA is obtained in the eluent with a pH lower than 4.5.

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: 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: This invention discloses a fibrinophilic urokinase complex which is a complex of urokinase with a urokinase inhibitor or tissue activator inhibitor and has a molecular weight of 97,500.+-.3,000 and a method for the production of this fibrinophilic urokinase complex from urine. The urokinase complex can be used, as a thrombolytic agent exhibiting an outstanding ability to dissolve thrombus.

Abstract: Applicant has isolated a Factor X.sub.a inhibiting substance from the salivary gland extract of the leech H.ghilianii. The extract is subjected to separation using both DEAE-cellulose and heparin-agarose chromatography resins eluting with an increasing salt gradient. The extract is subjected to affinity chromatography using Factor X.sub.a bound to an Affi-Gel-15 resin eluting with HEPES containing benzamidine. Reverse phase chromatography yields several purified peptides or proteins having Factor X.sub.a inhibiting activity. The properties and clinical compositions of these FX.sub.a inhibitors are described.

Abstract: A calcium-phosphate type hydroxyapatite which belongs to a series of hexagonal systems has unit lattice constants of 9.58.+-.0.08 .ANG. for the a axis and 7.00.+-.0.05 .ANG. for the c axis. Its Ca/P ratio is in the range of 1.50 to 1.90. It is for use as a column packing material for chromatographic separation of biopolymers. The hydroxyapatite is produced by firing it in the form of a gel or powder at a temperature of 400.degree. to 700.degree. C. As a gel the hydroxyapatite takes the form of a suspension or slurry. A powder is prepared by removing moisture from the hydroxyapatite in the gel form and then drying. Either gel or powder is fired by heating in the presence of oxygen or air at 500.degree. to 600.degree.0 C.

Abstract: Two components, trypsin and kallidinogenase, in human urine are concentrated simultaneously by allowing human urine at neutral pH, collecting bubbles thus formed to obtain the concentrate of the two components, adjusting the concentrate to weak acidity, contacting the acidified concentrate with chitosan to allow the two components to be adsorbed onto chitosan, eluting the components from the adsorbent with aqueous ammonia solution, and neutralizing and heating the eluate at about 60.degree. C. for about 10 hours to make the eluate virus-free, followed by separating the components from the eluate.

Abstract: A new procedure for isolating histamine N-methyltransferase in pure form has permitted the development of a new and highly sensitive radioenzymatic assay for histamine.

Abstract: A continuous industrial separation process for biopolymer extracts each individual single component from a stream of cell extract. The process can be set up as a mixed flow reactor or fluidized bed in a continuous operation under normal pressure to treat a volume of cell extract in a short period of time. The separated single biopolymer is continuously withdrawn from the process and be easily condensed into the desired concentration. A resin, which can be an ion exchanger or affinity adsorbent or dye ligand adsorbent or hydrophobic adsorbent or immunoadsorbent, is evenly suspended in a well mixed reactor to equilibrate with the biopolymers in the liquid phase. The fundamental separation scheme is successive adsorption and desorption of biopolymer from liquid. Biopolymers are isolated into a single stream or multistreams for one component or many individual components. Each stream contains only one high purity biopolymer.

Abstract: Deacetoxycephalosporin C synthetase is provided in purified form via chromatography of crude cell-free extracts over a weak anion exchange resin followed by gel filtration and hydroxylapatite chromatography, all carried out in the presence of glycerol, a C.sub.1 -C.sub.3 alkyl monohydric alcohol, e.g., ethanol, a sulfhydryl containing reducing agent, e.g., dithiothreitol, and ascorbate. The purified enzyme which possesses both expandase and hydroxylase activities can be further purified by chromatography over a strong anion exchange resin.

Abstract: A method for the purification of a culture filtrate resulting from the fermentation of an organism of the species Mucor miehei involves the selective adsorption of the microbial rennet present in the culture filtrate by a blue dye affinity ligand with subsequent elution of the adsorbed microbial rennet to provide it in a purified form.

Abstract: A chromatographic column and process for isolating or purifying steroid hormone or cell membrane receptors using the column are provided in which the column contains at least three resin layers between the inlet and the outlet ends of the column, the layer closest the inlet being a strong cationic exchange resin, the middle layer being a matrix containing a triazine dye that will bind proteases and proteins with dinucleotide fold conformations, and the layer closest the outlet being a weak anionic exchange resin, the ratio of the volumes of the resin layers being 1:2:1.

Abstract: A process for the recovery and purification of L-asparaginase from Erwinia chrysanthemi is disclosed. The process involves the preparation of cellular acetone powder extract followed by either an ion exchange and affinity chromatography purification steps or by affinity chromatography alone. The column eluent is then dialyzed to produce substantially pure L-asparaginase.

Abstract: Porous glass beads for filtation applications having a homogeneous metaloxane structure and comprising oxides of Si, Zr and optionally Ti and Al. A preferred method for making these beads comprises the steps of (a) providing a mother solution of Si and Zr alkoxides in a water soluble solvent, for instance a lower aliphatic alcohol, (b) providing a liquid dispersant phase in which solution (a) is dispersible and stirring this liquid phase sufficiently to cause (a) to be formed into droplets of substantially uniform size when added to (b), (c) adding (a) to (b) at a rate sufficient to provide said droplets and effecting the hydrolysis of the alkoxides contained therein with consecutive gelation of said droplets into corresponding hardened beads of condensed mixed Si and Zr hydroxides, and (d) separating said beads from the liquid phase and drying to achieve the desired porous mixed oxide structure for the beads.

Abstract: A batch system apparatus for collecting lysozyme from egg white by adsorption on an ion-exchange resin, which includes a lysozyme adsorption tank, equipped therein with a stirring mechanism for stirring an egg white solution and an ion-exchange resin, having a strainer provided in a lower portion thereof, having liquid discharge pipes connected thereto at a position below the strainer, preferably at the bottom of the tank, and at the side wall above the strainer at a height such that the resin when left at rest will not overflow, respectively, and further having a resin discharge pipe connected thereto at the lower portion of the side wall, above and in the vicinity of the strainer; a positive displacement pump connected to the liquid discharge pipes for transferring the egg white solution after removal of lysozyme to a reservoir tank; and a centrifugal pump connected to the resin discharge pipe for transferring the mixture of the resin and water to a separate tank, whereby collection by adsorption can be mad

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: The invention relates to ATP: polynucleotide adenylyltransferase enzyme and a method for its preparation comprising extraction from monocotyledonous plant tissue and chromatographic separation.

Abstract: Enzymes extracted from a prokaryotic .beta.-lactam producing microorganism are immobilized on a support for producing desacetoxycephalosporin and analogs thereof from L-.alpha.-aminoadipyl-L-cysteinyl-D-valine and analogs thereof. The enzymes are an epimerase, a cyclase and a ring expansion enzyme extracted preferably from S. clavuligerus, S. cattleya or S. lipmanii. The support is preferably a diethylaminoethyl ion exchange resin.

Abstract: A method is disclosed for the recovery of biological material from an aqueous solution comprising contacting a water-insoluble, particulate binder with a solution containing biological material to produce a water insoluble biological material/binder composition which may then be recovered. The aqueous solutions may then be recycled.

Abstract: The subject of the application is a process and a reagent for the differentiated determination of the bone and liver isoenzyme of alkaline phosphatase in a sample, whereby this is mixed with a lectin which is able to bind N-acetylglucosamine residues and is incubated. Thereafter, there is carried out a separation of the lectin-bound from the free isoenzyme portions and the alkaline phosphatase activity is determined in one or in both of the separated media.

Abstract: This invention relates to three enzymes, their isolation from the viscera of bivalves, e.g. the surf clam or cherrystone clam, their characterization and uses. The first two are carboxyl proteinases having molecular weights of about 77,200 and about 36,700 and display activity similar to mammalian D-cathepins. The third is a thiol proteinase having a molecular weight of about 17,400 and displays activity similar to mammalian B-cathepins. In addition to attaching various substrates, the enzymes coagulate cheese milk and tenderize meat.

Abstract: A method for treating a biological or pharmaceutical product to inactivate viruses and pyrogens therein comprising the steps of adsorbing said product onto a solid phase; treating the adsorbed product with a virus or pyrogen inactivating agent; separating the solid phase and quantitatively removing the residual inactivating agent therefrom; and recovering said product.

Abstract: Disclosed is a method for the purification of calf rennet which involves contacting the rennet with an affinity ligand, e.g. Cibacron Blue, to cause separation of the chymosin and pepsin contained in the rennet.

Abstract: The enzyme alkaline phosphatase having the following properties:(i)molecular weight: approximately 80 000 with 2 subunits of 40 000(iii)activator: Mg.sup.++inhibitor: EDTA(iii)thermal stability: 45.degree. C. or below(iv)optimum temperature: 35.degree.-45.degree. C.(v)pH stability: 6-11(vi)pH optimum: 10(vii)specific activity: 3000-5000 units/mg(viii)isoelectric range: between pH 5.0 and 6.0.An antibody-enzyme conjugate process for purifying alkaline phosphatase and a reagent test kit using the purified alkaline phosphatase are also disclosed.

Abstract: The present invention provides a process for the determination of N-carbamoylsarcosine, wherein a sample solution containing N-carbamoylsarcosine is reacted with N-carbamoylsarcosine-amidohydrolase to give sarcosine, which is then determined.The present invention also provides the enzyme N-carbamoylsarcosine-amidohydrolase, a process for obtaining it and a reagent containing it.

Abstract: Substantially pure rubber polymerase and analogues thereof are disclosed. Substantially pure rubber polymerase is isolated and purified from chemically stabilized Hevea brasiliensis latex. The analogues exhibit substantial homology to the Hevea brasiliensis polymerase or function in an enzymatically equivalent manner. Methods for the isolation and purification of a rubber polymerase are set forth. Use of substantially pure rubber polymerase to produce natural rubber or related copolymers is also disclosed.

Abstract: Alpha-1-antiprotease is highly purified from an impure solution thereof by a series of chromtographic procedures.

Abstract: The present invention relates to a process for the production of a purified glucose isomerase which comprises contacting an impure extract containing glucose isomerase and soluble impurities with a weakly basic ion exchange material known to adsorb glucose isomerase; adding a first salt solution of a concentration which removes unadsorbed and weakly adsorbed impurities, but not glucose isomerase; and adding a second, buffered salt solution which elutes the adsorbed glucose isomerase.

Abstract: Meat is tenderized by adding thereto a proteolytic enzyme obtained by culing the microorganism, Trichoderma reesei strain MCG 80. The enzyme is an aspartic acid protease with proteolytic properties similar to the animal protease, Cathepsin D. The enzyme acts selectively upon the myofibrillar proteins of meat producing a desirable uniform texture. Culturing of the microorganism in a medium containing glucose and lactose results in high enzyme yield.

Abstract: The present invention provides an efficient process for purifying the enzyme phenylethanolamine N-methyltransferase suitable for use in radioenzymatic assays of endogenous compounds.

Abstract: Disclosed is a process for purifying an enzyme contained in a solution such as cell extract liquor or fermentation culture liquor. The crude enzyme solution is brought into contact with either a strongly acidic cation exchange resin of high porous type or a strongly basic anion exchange resin of high porous type to adsorb the enzyme on the resin. An eluting agent is then passed through the resin to elute out the enzyme as a purified enzyme solution.

Abstract: The invention relates to a device useful when installed in a urinal for collection of urinary proteins. The adsorbent means includes a conduit for gravity feed passage of urine streams. The conduit has permeable layers in series containing a slow releasing antimicrobial agent and an adsorbent for urinary proteins such that for collection puropses wanted proteins are adsorbed without odor or loss due to bacterial degradation.

Abstract: Cyclase, epimerase and a ring expansion enzyme are isolated separately from a cell free extract of a prokaryotic beta-lactam producing organism to provide three separate and stable enzyme reagents for commercial production of cephalosporins from peptide precursors. Isolation is carried out by using ammonium sulfate, gel filtration and ion exchange chromatography. The enzymes may be immobilized on a suitable support and the production of cephalosporins may be carried out continuously.

Abstract: Aminoacylase is isolated from mammal kidneys by comminuting and homogenizing mammal kidney in water, centrifuging to form an aqueous extract, heating the aqueous extract at 60.degree. to 80.degree. C. for 5 to 15 minutes, centrifuging, adding a salt such as ammonium sulfate to the resultant supernatant, centrifuging to separate solids, dissolving the solids in water, dialyzing and recovering active aminoacylase from the dialyzed solution. The process produces aminoacylase with high specific activity and the process requires fewer purification steps. The aminoacylase obtained may be utilized directly without any subsequent purification to produce immonobilized aminoacylase. Immobilization can be carried out by covalent bonding of the aminoacylase to a partially hydrolyzed Akrilex P type acryl amide-N,N'-methylene-bis(arylamide) copolymer. The recovered aminoacylase can be subjected to two chromatographic purification steps to produce a very pure aminoacylase.

Abstract: In the process of eluting egg white lysozyme by contacting a weakly acidic cation exchange resin having egg white lysozyme adsorbed thereon with an eluting agent comprising a salt solution, an alkali agent is added to the eluate containing the weakly acidic cation exchange resin having egg white lysozyme adsorbed thereon whereby the egg white lysozyme can be eluted more effectively. From the eluate thus obtained, lysozyme can be collected in a high yield by an ordinary collecting method such as salting out.

Abstract: The enzymes, cyclase, epimerase and a ring expansion enzyme, are isolated separately from a cell-free extract of a prokaryotic beta-lactam producing organism. The isolated enzymes are used to produce unnatural cephalosporins from polypeptide precursors. Isolation is carried out by adding ammonium sulfate to 40% saturation to the cell-free extract to precipitate contaminating proteins, adding ammonium sulfate to 70% saturation to the resultant supernatant to precipitate the enzymes, suspending the precipitated enzymes in a buffer, separating epimerase from the suspension by gel filtration, and separating cyclase and the ring expansion enzyme from each other by ion exchange chromatography.