Journal of the Korea Academia-Industrial cooperation Society (한국산학기술학회논문지)

The Korea Academia-Industrial cooperation Society (한국산학기술학회)
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Production of antibodies for saxitoxin analysis and sensitivity analysis of anti-saxitoxin antiserum

삭시톡신 분석을 위한 항체의 제조 및 항-삭시톡신 항혈청의 민감도 분석

  • Received : 2012.09.05
  • Accepted : 2012.12.06
  • Published : 2012.12.31
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Abstract

The most essential but missing components to understand and use toxic substances from marine microalgae are developing the fast, easy and economical determining technology for detecting it. In this paper we produced the antibodies against saxitoxin (STX). Mariculture keyhole limpet hemocyanin (mcKLH) and ovalbumin (OVA) were used as carrier proteins. mcKLH-STX conjugates were injected into the peritonial cavity of BALB/c mouse for immunization. After bleeding from mouse, anti-STX antiserum was isolated. Indirect enzyme-linked immunosorbent assays (ELISA) was performed to determine antiserum titer using the microtiter plate coated with free STX and OVA-STX. A goat anti-mouse IgG-phosphatase conjugate was used as secondary antibody to enable chromogenic reaction. Reactions of anti-STX antiserum were very specific on the OVA-STX and free STX. Sensitivity of anti-STX antiserum on STX was very high and STX detection limit was to be 64.9 ng/kg for indirect ELISA.

해양미세조류 유래 독성물질에 대한 이해와 활용에 있어서 가장 중요하지만 간과되고 있는 부분은 독성물질을 검출할 수 있는 빠르고, 쉽고 경제적인 검출기술을 개발하는 것이다. 이 논문에서 우리는 삭시톡신(STX)에 대한 항체를 생산하였다. 헤모시아닌(mariculture keyhole limpet hemocyanin, mcKLH)과 오브알부민(ovalbumin, OVA)을 운반단백질로 사용하였다. 면역반응을 위해서 mcKLH-STX 결합체를 BALB/c 쥐에 복강주사하였다. 채혈 후 항-STX 항혈청을 분리하였다. 항혈청의 역가분석을 위하여 유리 STX와 OVA-STX로 코팅된 microtiter plate를 이용하여 간접 ELISA 실시하였다. 발색반응을 위한 이차항체로는 goat anti-mouse IgG-phosphatase conjugate가 사용되었다. 항-STX 항혈청은 OVA-STX와 유리 STX에 특이적으로 반응하였다. 항-STX 항혈청의 민감도는 매우 높았으며, STX를 위한 검출한계는 약 64.9 ng/kg이었다.

Keywords

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