서지주요정보
Studies on binding of clostridium botulinum type B toxin to rat brain synaptosomes and preparation of monoclonal antibodies to the toxin = Clostridium botulinum Type B 독소의 Rat brain synaptosome 에의 결합 성질 및 이 독소에 대한 단일 항체에 관한 연구
서명 / 저자 Studies on binding of clostridium botulinum type B toxin to rat brain synaptosomes and preparation of monoclonal antibodies to the toxin = Clostridium botulinum Type B 독소의 Rat brain synaptosome 에의 결합 성질 및 이 독소에 대한 단일 항체에 관한 연구 / Moon-Kook Park.
저자명 Park, Moon-Kook ; 박문국
발행사항 [대전 : 한국과학기술원, 1990].
Online Access 원문보기 원문인쇄

소장정보

등록번호

8001601

소장위치/청구기호

학술문화관(문화관) 보존서고

DBE 9008

휴대폰 전송

도서상태

이용가능

대출가능

반납예정일

초록정보

Clostridium botulinum type B toxin was purified by column chromatography steps with DEAE-Sephadex and Sephacryl S-300. Heavy chain and light chain of type B-toxin were prepared by elution from SDS-polyacrylamide gels. The toxin and subunits were labeled with [$^{125}I$]iodine. To clarify the role of each subunit in binding of botulinum toxin to target cells, binding of labeled toxin and subunits to rat brain synaptosomes was studied. When labeled toxin or subunit was incubated with various amounts of rat brain synaptosomes, light chain did not bind to synaptosomes. In contrast, specific binding of toxin and heavy chain was observed and there was no significant difference in the amounts of toxin and heavy chain bound. When competitive effects of unlabeled toxin and heavy chain on the binding of labeled toxin and heavy chain were examined, binding of labeled toxin and heavy chain to synaptosomes was inhibited to a similar degree by unlabeled toxin. Unlabeled heavy chain showed similar inhibitory effects as unlabeled toxin. The half-inhibition concentrations of unlabeled heavy chain for the bindings of labeled toxin and heavy chain were similar to those of unlabeled toxin. These results indicate that the heavy chain has similar binding properties to synaptosomes as the intact toxin and may also suggest that the heavy chain alone is involved in the binding of botulinum type B toxin to synaptosomes. BALB/c mice were immunized with type B toxoid or heavy chain, and spleen cells from immunized mice were fused with Sp2/O-Ag14 myeloma cells. Antibody-secreting hybridomas were screened by enzyme-linked immunosorbent assay, and cloned by limiting dilution. Four cell lines were established. The four monoclonal antibodies produced by them were characterized (Sub) Classes were determined by double immunodiffusion tests. ELISA and Immunoblotting tests with heavy and light chain showed that the all recognize the heavy chain of type B toxin. In the neutraliation tests with mice, antibody B3 had much higher neutralizing activity than the other three. When antibodies were examined for the effects on binding of labeled toxin and heavy chain to synaptosomes antibody B3 inhibited the binding effectively and the others had little inhibitory effects. This result indicates that antibody B3 blocks the 'binding site' in the heavy chain and thus prevents binding of the toxin to target cell which results in neutralizing toxic activity of the toxin whereas antibodies B1, B2, and B4 recognize some other sites in the heavy chain rather than the 'binding site.'

서지기타정보

서지기타정보
청구기호 {DBE 9008
형태사항 x, 99 p. : 삽도 ; 26 cm
언어 영어
일반주기 저자명의 한글표기 : 박문국
지도교수의 영문표기 : Kyu-Hwan Yang
지도교수의 한글표기 : 양규환
학위논문 학위논문(박사) - 한국과학기술원 : 생물공학과,
서지주기 Reference : p. 90-99
주제 Botulinum toxin.
Bacterial toxins.
Monoclonal antibodies.
세균 독소. --과학기술용어시소러스
보툴리누스 독소. --과학기술용어시소러스
단일 클론성 항체. --과학기술용어시소러스
Proteins --Separation.
단백질 분리
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