| Project/Area Number |
62580140
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
物質生物化学
|
|---|
| Research Institution | Tokyo Metropolitan Institute of Medical Science |
|---|
Principal Investigator |
INAGAKI Fuyuhiko Tokyo Metropolitan Institute of Medical Science, Section Chief, 生理活性物質研究部門 (70011757)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
KOHDA Daisuke Tokyo Metropolitan Institute of Medical Science, Researcher, 生理活性物質研究部門, 研究員 (80186618)
KASE Ryouichi Tokyo Metropolitan Institute of Medical Science, Researcher, 生理活性物質研究部門, 研究員 (20150203)
KITAGAWA Hisayo Tokyo Metropolitan Institute of Medical Science, Researcher, 循環器病研究部門, 主任研究員 (70124451)
|
|---|
| Project Period (FY) |
1987 – 1988
|
| Project Status |
Completed (Fiscal Year 1988)
|
| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1988: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1987: ¥1,200,000 (Direct Cost: ¥1,200,000)
|
|---|
| Keywords | alpha-Bungarotoxin / Monoclonal antibody / hybridoma / Fab fragment / epitope Deuterium labell / Fab / エピトープ / α-ブンガロトキシン / ヘビ神経毒タンパク / 核磁気共鳴 |
|---|
| Research Abstract |
alpha-bungarotoxin was purified from the venom of Bungarus multicinctus. Immunization of mice was achieved by injecting subcutaneously native toxin at two week intervals using the incomplete Freunds' adjuvant into BALB c mice. After the conventional methods, nine hybridomas producing monoclonal antibodies specific to alpha-bungarotoxin were obtained. Large quantity of monoclonal antibody MaB-2311 produced by one of these hybridoma was purified from the ascites fluid of BALB c mice. MaB-2311 inhibited the lethal effect of alpha-bungarotoxin in vivo assay, suggesting that this monoclonal antibody binds to the region specific to neurotoxic activity. Cross reactivity of MaB-2311 was studied using several long neurotoxins, including Toxin B, Toxin III and LS III. Compared with the cross reactivity of these toxins and MaB-2311, and homology of the primary sequences, an antigenic determinat for MaB-2311 was determind to be Ser34-Ser35-Arg36-Gly37-Lys38. The peptide fragment containing this sequence also reacts with MaB-2311, supporting again that the region involving this sequence is an antigenic determinat. Since Arg36 is an essential residue for neurotoxicity, it is reasonable that this monoclonal antibody neutralizes the neurotoxicity of alpha-bungarotoxin. The MaB-2311 producing hybridoma was subsequently cultured in serum free medium so that deuterium labelled amino acids were incorporated into MaB-2311. NMR studies to investigate the molecular recognition of alpha-bungarotoxin and MaB-2311 are now in progress using the deuterium labelled monoclonal antibody and the peptide fragment.
|
