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    HIGH-MOLECULAR LINK BETWEEN METHOTREXATE DERIVATIVE AND PYRAN COPOLYMER AND ITS PRODUCTION
    2.
    发明专利
    HIGH-MOLECULAR LINK BETWEEN METHOTREXATE DERIVATIVE AND PYRAN COPOLYMER AND ITS PRODUCTION 失效

    公开(公告)号：JPH07173080A

    公开(公告)日：1995-07-11

    申请号：JP13287393

    申请日：1993-05-11

    Applicant: AGENCY IND SCIENCE TECHN

    Inventor： HIRANO TAKASHI ,  TODOROKI TAKESHI ,  OHASHI SHINICHI ,  KOKUBU TOMOKUNI ,  TANAKA HIDEAKI

    IPC: A61K31/505 ,  A61K47/48 ,  A61P35/00

    Abstract: PURPOSE:To obtain the subject new low toxic high-molecular link or a salt thereof having excellent carcinostatic activity. CONSTITUTION:This high-molecular link is made up of 10-500 recurring units of formula I and formula II (R is a methotrexate residue of formula III or IV; (n) is 1-12) with the molar ratio of formula I to formula II of (0:100) to (90:10). The high-molecular link is obtained by the following processes: methotrexate is reacted with an alkyldiamine to synthesize and isolate a methotrexate derivative, which is, in turn, reacted with a pyran copolymer in a water-soluble organic solvent. The high-molecular link gradually releases the methotrexate derivative as a carcinostatic substance in the serum, having markedly excellent carcinostatic activity as compared to the single use of methotrexate, owing to the synergistic effect between carcinostatic methotrexate and carcinostatic pyran copolymer.

    ENZYME IMMUNOASSAY METHOD FOR LEUCINE ENKEPHALINE
    8.
    发明专利
    ENZYME IMMUNOASSAY METHOD FOR LEUCINE ENKEPHALINE 失效

    公开(公告)号：JPS63111465A

    公开(公告)日：1988-05-16

    申请号：JP25719086

    申请日：1986-10-29

    Applicant: AGENCY IND SCIENCE TECHN

    Inventor： KOKUBU TOMOKUNI ,  IWAKURA MASAHIRO ,  OHASHI SHINICHI ,  TSUDA KEISHIRO

    IPC: C12N15/02 ,  C12N15/00 ,  C12P21/00 ,  C12Q1/00 ,  G01N33/53 ,  G01N33/535 ,  G01N33/577

    Abstract: PURPOSE:To improve measurement sensitivity and reproducibility by utilizing the fused protein genetically bonded with labeling enzyme and antigen by a gene manipulation as a labeling antibody. CONSTITUTION:The coliform bacilli contg. the plasmid having the gene coding the dihydro folic acid reductase-leucine enkephalline fused protein (hereafter expressed as DHFR-L-Enk) are prepd. by the gene manipulation and are subjected to fractionating, refining and purifying after liquid culture to obtain the DHFR-L-Enk exhibiting a uniform band in SDS electrophoresis. Said liquid is added together with an antirabbit leucine enkephalline (hereafter expressed as L-Enk) antibody to cause reaction, by which a soln. for measuring enzyme activity is obtd. A DHFR substrate soln. is then added thereto to initiate reaction. A change in the absorption at 340nm wavelength is measured by a spectrophotometer, by which L-Enk is measured.

    PRODUCTION OF IMMOBILIZED ENZYME
    9.
    发明专利
    PRODUCTION OF IMMOBILIZED ENZYME 失效

    公开(公告)号：JPH0833485A

    公开(公告)日：1996-02-06

    申请号：JP32954091

    申请日：1991-11-18

    Applicant: AGENCY IND SCIENCE TECHN

    Inventor： IWAKURA MASAHIRO ,  KOKUBU TOMOKUNI ,  OHASHI SHINICHI

    IPC: C12N11/00 ,  C12N9/06

    Abstract: PURPOSE:To obtain a highly active immobilized enzyme for bioreactor, etc., in high efficiency by introducing an amino acid sequence having a cysteine residue at the terminal to the carboxy-terminal of an enzyme protein and immobilizing the modified protein to a carrier through an SH group. CONSTITUTION:This immobilized enzyme having high uniformity and activity and useful as an element of a bioreactor, etc., is produced by constructing an expression vector coding a modified enzyme protein obtained by introducing a short amino acid sequence to the carboxy terminal of an enzyme protein (e.g. dihydrofolate reductase) to form cysteine residue on the carboxy terminal of the protein, introducing the expression vector into a host cell, culturing the transformed cell to effect the expression of the gene and obtain a modified enzyme protein having an amino acid sequence composed of several number of amino acid residues having a cysteine residue terminal on the carboxy terminal of the enzyme protein and bonding the modified protein to an immobilization carrier through a mercapto group of the cysteine residue at the carboxy terminal.

    10.
    发明专利
    失效

    公开(公告)号：JPH05339295A

    公开(公告)日：1993-12-21

    申请号：JP34867791

    申请日：1991-12-05

    Applicant: AGENCY IND SCIENCE TECHN

    Inventor： IWAKURA MASAHIRO ,  KOKUBU TOMOKUNI ,  OHASHI SHINICHI

    IPC: C07K16/00 ,  C07K14/00 ,  C07K19/00 ,  C12N15/09 ,  C12N15/62 ,  C12P21/02 ,  C12R1/19 ,  C07K15/14

    Abstract: PURPOSE:To produce an antibody by bonding the objective peptide to the carboxyl terminal of dihydrofolic acid reductase(DHFR) and using the produced fused protein as an immunizing antibody. CONSTITUTION:The objective process for the preparation of an antibody against a peptide is composed of a process for producing a DHFR fused protein having the objective peptide on the carboxyl terminal side by genetic engineering technique, a process for separating the objective fused protein from the host microorganism and purifying the protein and a process for immunizing an animal using the objective highly purified fused protein.

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