The Korean Journal of Pesticide Science (농약과학회지)
- Volume 5 Issue 2
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- Pages.1-12
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- 2001
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- 1226-6183(pISSN)
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- 2287-2051(eISSN)
ELISA Development for the residue of the organophosphorus insecticide acephate
ELISA에 의한 유기인계 살충제 Acephate 잔류물 분석법 개발
- Lee, Jae-Koo (Department of Agricultural Chemistry, College of Agriculture, Chungbuk National University) ;
- Ahn, Ki-Chang (Department of Agricultural Chemistry, College of Agriculture, Chungbuk National University) ;
- Stoutamire, Donald W. (Departmnent of Entomology and Cancer Research Center, University of California) ;
- Gee, Shirley J. (Departmnent of Entomology and Cancer Research Center, University of California) ;
- Hammock, Bruce D. (Departmnent of Entomology and Cancer Research Center, University of California)
- Published : 2001.06.30
Abstract
A competitive indirect enzyme-linked immunosorbent assay (ci ELISA) for the organophosphorus insecticide acephate, O,S-dimethyl acetylphosphoramidothioate, was developed using a polyclonal antibody. Three different haptens mimicking the analyze and containing hexanoic acid moiety as a linker were synthesized, and then conjugated with the carrier proteins bovine serum albumin and keyhole limpet hemocyanin by the N-hydroxysuccinimide active ester method. Polyclonal antibodies raised against hapten-KLH conjugates in rabbits and the hapten-BSA conjugates as coating antigens were screened and selected for the assay in the homologous and/or heterologous ELISA system. The effects of various assay conditions, including blocking reagents, detergent content, organic solvents, pH, and preincubation of tile mixture of the polyclonal antibody and the analyze on the sensitivity were evaluated. The
다클론항체를 이용하여 환경 시료중 유기인계 살충제 acephate를 분석할 수 있는 경합적 간접효소면역측정법(competitive indirect ELISA)을 개발하기 위하여 분석물과 화학구조가 유사하며 hexanoic acid를 지닌 3종의 hapten을 합성하였다. 이들 hapten을 active ester 방법으로 담체단백질인 keyhole limpet hemocyanin과 접합시켜 면역원으로 사용하있고, bovine serum albumin과 접합시켜 homologous 혹은 heterologous ELISA를 위한 코팅항원으로 사용하였다. 생산된 항체들과 코팅항원을 최종선발하여 acephate 잔류분석을 위한 ELISA를 위하여 최적화하였다. 단백질의 농도가