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Monoclonal Antibodies to Spirosin of Yersinia enterocolitica and Analysis of the Localization of Spirosome by Use of Them
Author(s) -
Yamato Masayuki,
Takahashi Yuko,
Tomotake Hiroyuki,
Ota Fusao,
Hirota Katsuhiko,
Yamaguchi Kazuo
Publication year - 1994
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.1994.tb01762.x
Subject(s) - monoclonal antibody , yersinia enterocolitica , biology , microbiology and biotechnology , antibody , western blot , proteolysis , yersinia , blot , spleen , protease , immunochemistry , clone (java method) , cell fractionation , antiserum , cytoplasm , bacteria , biochemistry , immunology , dna , enzyme , genetics , gene
Two hybridomas producing monoclonal antibodies (MAbs) were prepared by fusing myeloma cells (Sp2/0‐Ag14) with mouse spleen cells immunized with purified spirosin from Yersinia enterocolitica SYT‐11–72 (YE72). The antibodies produced by them were designated MAbs‐S5 and S27. They were IgG 2a and IgG 1 , respectively, both with χ light chains. MAbs‐S5 and S27 reacted specifically with spirosin from YE72. On Western blotting after limited proteolysis with Staphylococcus aureus V8 protease, YE72 spirosin revealed peptide fragments of 35 and 37 kDa reacting markedly with MAb‐S5, which suggested the presence of an antigenic determinant on these fragments. By cellular fractionation of YE72 and subsequent EIA and Western blot analysis, spirosome was shown to be present in the cytoplasm of YE72.