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Generation of Polyclonal Antibody against μ-Conotoxin GIIIA Using an Immunogen of [Cys5]μ-Conotoxin GIIIA Site-Specifically Conjugated with Bovine Serum Albumin

2002; Elsevier BV; Volume: 290; Issue: 3 Linguagem: Inglês

10.1006/bbrc.2001.6276

1090-2104

Mitsuhiro Nakamura, Yuichi Oba, Tatsuya Mori, Kazuki Sato, Yukisato Ishida, Tsukasa Matsuda, Hideshi Nakamura,

Receptor Mechanisms and Signaling

μ-Conotoxin GIIIA, one of the strong peptide toxins in the cone shell, preferentially blocks the skeletal muscle-type sodium channels in vertebrates. The toxicity of μ-conotoxin GIIIA is nearly equal to that of tetrodotoxin. The generation of an antibody for the native toxins is analytically useful, but practically difficult due to its high toxicity to animals. In this study, we generated the polyclonal antibody for μ-conotoxin GIIIA using a specific conjugation method in which the immunogen was detoxified while retaining the active-site structure for the sodium channels. ELISA analysis showed that the generated antibody recognized the native toxin folded with three disulfide bridges, but not the linear one. Furthermore, the physiologically active mutants of GIIIA were recognized while the inactive mutants were not, suggesting that the newly generated antibody can selectively recognize the physiologically active toxins. These methods for generating an antibody against peptide toxins will be applicable to other peptide toxins.