한국식품위생안전성학회지 Vol.32 No.1 (p.75-81)

고등어 어육 중 열안정성 단백질에 특이한 단클론성 항체 개발과 특성 확인

Production of A Monoclonal Antibody (MAb) Against a Thermal Stable-Soluble Protein in Mackerel and Confirmation of the Properties for the MAb
키워드 :
Mackerel,monoclonal antibody,ELISA,western blot

목차

Materials and Methods
시료 및 실험재료
  TSSP의 추출
  Buffer에 따른 TSSP 추출효율 확인
  면역 항원 준비
  단클론성 항체 생산
  단클론성 항체 특성 확인
Results and Discussion
  고등어 어육 중 TSSP의 확인
  TSSP 추출법 최적화
  단클론성 항체 생산 및 반응성 확인
  생산된 항체를 이용한 효소면역분석법과 western blot의검출한계 확인
Acknowledgement
국문요약
References

초록

For people who have a food allergy the only way to manage the allergy is to avoid the food allergen. The mackerel is one of the major food allergens, but no immunoassay for the rapid and simple detection of mackerel has been reported. The objectives of this study are to develop and characterize monoclonal antibodies (MAbs) specific to mackerel using thermal stable-soluble proteins (TSSP) as an immunogen and to characterize the MAbs by indirect enzyme-linked immunosorbent assay (iELISA). The mice immunized with mackerel TSSP and showing high titer were used for cell fusion and cloning. The characterization of MAbs produced from hybridoma cells obtained was confirmed by indirect ELISA and western blot. Four MAbs were confirmed to be specific to mackerel without crossreaction to other marine products and livestock products in the both methods. The iELISA and western blot based on the MAbs can sensitively detect 1% mackerel protein in other marine products. These results support that immunochemical methods based on the MAb produced could be used as rapid means to detect low levels of mackerel and to identify mackerel adulterated in food.