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Production of Monoclonal Antibody to 2′,3′‐Cyclic Nucleotide 3′‐Phosphodiesterase from Bovine Cerebral White Matter
Author(s) -
Fujishiro Masatoshi,
Kohsaka Shinichi,
Nagaike Kazuhiro,
Tsukada Yasuzo
Publication year - 1986
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1986.tb02849.x
Subject(s) - monoclonal antibody , microbiology and biotechnology , biology , myelin , immunohistochemistry , antibody , blot , oligodendrocyte , white matter , central nervous system , biochemistry , immunology , endocrinology , gene , medicine , radiology , magnetic resonance imaging
Lewis rats were immunized with partially purified 2′,3′‐cyclic nucleotide 3′‐phosphodiesterase (CNPase) from bovine cerebral white matter and the spleen cells were fused with cell of a mouse myeloma cell line (SP‐2). The production of monoclonal antibody was detected by (1) enzyme‐linked immunoadsorbent assay, (2) immunohistochemical staining of bovine cerebrum, (3) Western blotting analysis, and (4) CNPase binding assay. Monoclonal antibody that specifically binds CNPase molecules was obtained. However, the antibody did not suppress the enzyme activity. Western blotting analysis demonstrated that the monoclonal antibody binds both CNa (Wl a ) and CNb (Wl b ). The monoclonal antibody was identified as being of the IgG 2c subclass. Immunohistochemical examination revealed that the myelin sheath in the CNS was heavily stained with the monoclonal antibody in several species (bovine, mouse, rat, and human). In contrast, peripheral nervous system myelin was not stained even in bovine tissue. These results suggest that the monoclonal antibody obtained in the present study specifically recognizes the CNPase molecules in the CNS.