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Please use this identifier to cite or link to this item: http://hdl.handle.net/20.500.12001/3462

 
Title :[原著]An improved method for the detection of IgE FE-3 by ELISA using monoclonal anti-IgE FE-3 antibody
Authors :Nakasone,Toshiyuki
Hanashiro, Kazuhiko
Nakamura, Mariko
Sunakawa, Hajime
Kosugi, Tadayoshi
Authors affiliation :1st Department of Physiology, School of Medicine, University of the Ryukyus,
Department of Oral and Maxillofacial Surgery, School of Medicine, University of the Ryukyus,
Issue Date :2001
Publisher :琉球医学会
Type Local :雑誌掲載論文
Type (NII) :Journal Article
ISSN :1346888X
0289-1530
Abstract :A hybridoma producing monoclonal antibody against monoclonal DNP-specific rat IgE (IgE FE-3) was generated by fusion of P3-X63-Ag8-Ul (P3U1) mouse myeloma and spleen cells from BALB/c mice immunized with IgE FE-3. Five hybridoma clones were obtained. Three clones of them had a high reactivity against IgE FE-3 and a low reactivity against the L-chain of IgE FE-3 on ELISA analysis for the first screening of the hybridoma. The binding of monoclonal antibodies produced from clone BB-9 to IgE FE-3 revealed the strongest inhibition among three monoclonal antibodies on pretreatment with polyclonal rabbit anti-IgE FE-3 antibodies. It was clarified that monoclonal antibody BB-9 reacted specifically with the heavy chain of IgE FE-3 by Western blotting analysis. Furthermore, the reactivity of the monoclonal antibody BB-9 to IgE FE-3 was higher than that of MARE-1, which is a monoclonal antibody against rat myeloma IgE. The monoclonal antibody BB-9 was therefore employed for IgE-capture ELISA to determine the concentration of IgE FE-3. No significant alterations of color development occurred when igE FE-3 was concomitantly incubated with large excesses of IgG antibodies and rat myeloma IgE. Our data indicated that IgE-capture ELISA employing the monoclonal antibody BB-9 had a higher sensitivity for the determination of IgE FE-3 than did that using polyclonal rabbit anti-IgE FE-3 antibodies.
Rights :琉球医学会
URI :http://hdl.handle.net/20.500.12001/3462
Citation :琉球医学会誌 = Ryukyu Medical Journal Vol.20 no.3 p.129 -135
Appears in Collections:Ryukyu Medical Journal

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