公开(公告)号：JPH0564583A

公开(公告)日：1993-03-19

申请号：JP2389992

申请日：1992-02-10

Applicant: TORAY INDUSTRIES

Inventor： OSHIHARA WATARU ,  KOJIMA SHUNJI ,  NAKAMURA HARUJI

IPC: C12N9/02 ,  C12Q1/66 ,  C12Q1/68 ,  G01N21/76 ,  G01N33/532 ,  G01N33/535

Abstract: PURPOSE:To provide the subject modified Cypridina-derived enzyme bonded to a bioactive substance such as an antibody, an enzyme, a hormone, a toxin or a nucleic acid, excellent in stability, capable of application to various kinds of bioluminescence analysis and suitable for enzyme immunoassay measurement method, DNA probe method, etc. CONSTITUTION:A manifested vector pGL1 obtained by inserting Cypridina luciferase cDNA into the downstream of GAL1 promoter is introduced into an yeast by the protoplast method to obtain a transformant. The resultant transformant is subjected to shake culture in a culture medium at 30 deg.C for 2 day and the obtained cultured solution is added to a fermentation tank. After culture at 30 deg.C overnight, the supernatant of the cultured material is collected by centrifugation and a crude enzyme solution is obtained. The crude enzyme solution is then purified by gel filtration and the resultant purified luciferase is bonded to a low-molecular bioactive substance such as an antigen, a hapten, a hormone, an enzyme substrate, a receptor, a sugar chain or a coenzyme or a high-molecular bioactive substance such as an antibody, an enzyme, a hormone, a toxin or a nucleic acid, thus obtaining the objective modified luciferase excellent in stability.

公开(公告)号：JPS63129984A

公开(公告)日：1988-06-02

申请号：JP27669686

申请日：1986-11-21

Applicant: TORAY INDUSTRIES

Inventor： YANAI AKIRA ,  NAKAMURA NAOKO ,  OSHIHARA WATARU

IPC: C12N15/09 ,  C12N9/90 ,  C12N15/00 ,  C12R1/19

Abstract: PURPOSE:To improve the productivity of a microbial strain capable of producing alpha-amino-epsilon-caprolactam (ACL), by producing an ACL racemase having a specific amino acid sequence and a DNA sequence coding said racemase. CONSTITUTION:A gene DNA coding alpha-amino-epsilon-caprolactam (ACL) racemase is produced from a chromosome DNA of Achromobacter oobae (FERM P-776) or from a composite plasmid DNN36. A recombinant E.coli transformed with a composite plasmid DNN40 having structural gene of Achromobacter oobae or enzyme can be used as a bacterial strain capable of producing ACL racemase. The objective ACL racemase can be easily extracted from the recombinant E.coli and purified.

公开(公告)号：JPH08154699A

公开(公告)日：1996-06-18

申请号：JP29843294

申请日：1994-12-01

Applicant: TORAY INDUSTRIES

Inventor： MURAO YASUO ,  KONO HAJIME ,  OSHIHARA WATARU

IPC: C12Q1/66

Abstract: PURPOSE: To obtain the subject solution, containing a cypridinoid luciferin and an acidic solute, capable of enhancing the stability of an aqueous solution of the cypridinoid luciferin, improved in luminous efficiency and stability of measured values and useful for a high-sensitivity sensing system, etc., for an enzyme immunoassay. CONSTITUTION: A cypridinoid luciferin and an acidic salute such as formic, propionic, malic, tartaric, ethylenediaminetetraacetic or citric acid are dissolved in a 10mM phosphoric acid buffer physiological saline solution, etc., at pH

公开(公告)号：JPH0798316A

公开(公告)日：1995-04-11

申请号：JP24307693

申请日：1993-09-29

Applicant: TORAY INDUSTRIES

Inventor： OSHIHARA WATARU ,  KOJIMA SHUNJI ,  MASUKO SANAE

IPC: G01N33/532 ,  G01N33/543 ,  G01N33/574 ,  G01N33/577

Abstract: PURPOSE:To quickly measure the enzyme immunity of cancer-related antigen by detecting the luminescence of a trace constituent at the initial stage of reaction by sandwich immunoassay with bioluminescence enzyme cypridinacea luciferase. CONSTITUTION:The cypridinacea luciferase to be used is a protein having amino acid sequence, and the amino acid sequence may be partially substituted, missed, or inserted as far as the similar biological activity is held. When this marker enzyme is used, the trace quantity of 1-10ng is sufficient for one test as compared with 100ng or above of the marker enzyme required in the past. The luminescence based on the generated complex quantity immediately after the reaction before the antigen-antibody reaction reaches the saturated state can be detected, thus the enzyme immunity of a sample to be measured can be measured within 30min, preferably within 10 min, after it is added.

公开(公告)号：JPH0712817A

公开(公告)日：1995-01-17

申请号：JP15182893

申请日：1993-06-23

Applicant: TORAY INDUSTRIES

Inventor： OSHIHARA WATARU ,  HOSAKA SUKIKO

IPC: C12Q1/28 ,  G01N33/53 ,  G01N33/543 ,  G01N33/576

Abstract: PURPOSE:To quickly detect an antibody to be measured with high sensitivity without causing any environmental problem by separating the combined product of an immobilizing antigen and the antibody to be measured from a sample after immobilizing the antibody to be measured with the immobilizing antigen and measuring the amount of the remaining immobilizing antigen. CONSTITUTION:An immobilizing antigen prepared by immobilizing an antigen which specifically reacts to an antibody to be measured on an immobilizing carrier is made to react to a sample containing the antibody to be measured. Then the combined product of the immobilizing antigen and antibody to be measured is separated from the sample and cleaned. Thereafter, the immobilizing antigen remaining in the separated immobilizing carrier is made to react to a labeled antibody which specifically reacts to the antigen. After reaction, the amount of the label contained in the combined product immobilized on the immobilizing carrier is measured. From the measured amount, the amount of the antibody to be measured in the sample against the antigen can be detected indirectly.

公开(公告)号：JPH05113443A

公开(公告)日：1993-05-07

申请号：JP9973792

申请日：1992-04-20

Applicant: TORAY INDUSTRIES

Inventor： OKUDAIRA HIROICHI ,  OSHIHARA WATARU ,  HOSAKA SUKIKO

IPC: G01N33/535 ,  G01N33/543

Abstract: PURPOSE:To measure the quantities of specific antibodies in many items quickly at the same time so that the using amount of serum is small by using luciferase derived from a sea firefly as a label enzyme in the immunity measuring method for detecting a complex that is fixed to a solid carrier. CONSTITUTION:As a label enzyme, the luciferase derived from a sea firefly is used. The producing method of the sea-firefly luciferase can be any method. The luciferase is refined as required from the enzyme liquid containing the obtained luciferase activity. As the bonding method of an antibody and the lucifrase derived from the sea firefly, a bivalent reagent having the reactive group in the same reacting property or the different reacting property at both ends with a spacer in-between is used, and the complex of protein and protein is formed. As the reactive group of the bivalent reagent, e.g. n- hydroxysuccinimide or the like, which reacts with an amino group on the luciferase, can be listed. The antibody, which is used as the label antibody is appropriately selected for the material to be measured such as a human body and a mouse.

公开(公告)号：JPH04339260A

公开(公告)日：1992-11-26

申请号：JP28977691

申请日：1991-11-06

Applicant: TORAY INDUSTRIES

Inventor： OSHIHARA WATARU ,  KOJIMA SHUNJI

IPC: G01N33/535 ,  G01N33/543

Abstract: PURPOSE:To rapidly perform highly sensitive enzyme immunoassay by using luciferase occurring from a sea firefly as labelling enzyme. CONSTITUTION:Sea firefly luciferase to be used is protein having an amino acid sequence and, for example, it is collected from the natural world or prepared from an artifically bred sea firefly or a host cell other than the sea firefly by a gene recombination technique or cell culture technique and, if necessary, said liciferase is purified from an anzyme solution having luciferase activity. This luciferase is modified by a functional group or molecule of every kind to form a labelled antibody and, when this antibody is used in enzyme immunoassay, luminescence based on a very small amount of a component at the beginning of reaction when antigen-antibody reaction is not sufficiently advanced can be detected.

公开(公告)号：JPS62236487A

公开(公告)日：1987-10-16

申请号：JP8046986

申请日：1986-04-08

Applicant: TORAY INDUSTRIES

Inventor： YANAI AKIRA ,  NAKAMURA NAOKO ,  OSHIHARA WATARU

IPC: C12N15/09 ,  C07H21/04 ,  C12N1/20 ,  C12N9/90 ,  C12N15/00 ,  C12P13/08 ,  C12R1/19

Abstract: PURPOSE:To enable efficient production of enzyme for L-lysine production, by linking a gene coding alpha-amino-epsilon-caprolactam racemase to downstream of tac promoter. CONSTITUTION:A structural gene of alpha-amino-epsilon-caprolactam (ACL) racemase has been succeeded in linking to the downstream of tac promoter known as a strong promoter. It can be achieved e.g. by linking (A) an N-terminal of a structural gene of plasmid pNN38 produced by shortening plasmid pNN36 having a gene coding ACL racemase to (B) an EcoRI site of the downstream of tac promoter of plasmid pKKDELTANP obtained by shortening a plasmid pKK233-3 having tac promoter interposing (C) a start codon ATG between (A) and (B).

公开(公告)号：JPS60244289A

公开(公告)日：1985-12-04

申请号：JP9859084

申请日：1984-05-18

Applicant: TORAY INDUSTRIES

Inventor： YANAI AKIRA ,  NAKAMURA NAOKO ,  OSHIHARA WATARU

IPC: C12N15/09 ,  C07H21/04 ,  C12N9/90 ,  C12R1/19

Abstract: PURPOSE:To obtain a plasmid having cDNA of alpha-amino-epsilon-caprolactam (ACL), by expressing a gene in lysine requiring coliform bacilli, and converting the D-ACL into lysine in the presence of ACL hydrolase, and assimilating and multiplying the resultant transformant. CONSTITUTION:DNA fragments of a microorganism capable of producing alpha- amino-epsilon-caprolactam (ACL), e.g. Acromobacter obae, are integrated into a vector pBR322, and the resultant plasmid is used to transform lysine requiring variant strain of coliform bacilli. The resultant transformer is then selectively cultivated in a minimum essential culture medium, containing ACL hydrolase and D-ACL, and supplementing the nutritional requirement other than lysine to obtain a transformant having the ACL activity. A plasmid extracted from the transformant is then digested with EcoRI, and the resultant DNA fragments are integrated into a plasmid pACYC184 to give the aimed plasmid pNN32 containing the gene coding the ACL.

公开(公告)号：JP2000005301A

公开(公告)日：2000-01-11

申请号：JP16975498

申请日：1998-06-17

Applicant: TORAY INDUSTRIES

Inventor： SUGIZAKI HIROAKI ,  OSHIHARA WATARU

IPC: A61M1/14 ,  B01D61/30

Abstract: PROBLEM TO BE SOLVED: To prohibit the contact of a dialyzate with the sealing portion in a coupler for a blood dialyzer and to prevent the intrusion of the toxin derived from microorganisms into the dialyzate by installing a dialyzate tube which is insertable into the joint tube part of a dialyzate nozzle inserted in the coupler inside the coupler. SOLUTION: When the joint tube part of the dialyzate nozzle is inserted into the joint tube part 22 of the coupler, an O-ring 12 for sealing within the coupler is pressed by a ball locking mechanism to achieve liquid-tightness. Namely, a sleeve 14 is moved to the position of a stopper 20 by the elastic force of a coil spring 16, by which a ball 18 is locked and the liquid-tightness is maintained. In such a case, the dialyzate tube 24 consisting of a synthetic resin exhibiting rubber-like elasticity having an outside diameter to allow the insertion into the inside wall of the joint tube part of the dialyzate nozzle is installed into the coupler. As a result, the contact of the dialyzate with the sealing portion in the coupler is prohibited and the propagation of the microorganisms and the outflow of the toxin derived from the microorganisms to the dialyzate hose side through a juncture are prevented.