옥수수중 Deoxynivalenol의 검출을 위한 효소면역측정법의 개발

  • Lee, Hyang-Burm ;
  • Shon, Dong-Hwa ;
  • Kosaka, Kunio ;
  • Ueno, Yoshio
  • 이향범 (한국식품개발연구원 이화학연구부) ;
  • 손동화 (한국식품개발연구원 이화학연구부) ;
  • 코사카 쿠니오 (일본이과대학 약학부) ;
  • 우에노 요시오 (일본이과대학 약학부)
  • Published : 1997.08.01

Abstract

In order to develop an enzyme-linked immunosorbent assay (ELISA) for deoxynivalenol (DON) in com, we produced a specific monocl- onal antibody and established ELISA conditions. After the spleen cells from mice immunized with DON-bovine serum albumin conjugate were fused with S$_{p}$2/0 myeloma cells, a hybridoma cell 3G7 producing anti-DON antibody was screened by ELISA. From the standard curve of competitive direct ELISA (cdELISA) using 3G7 monoclonal antibody and DON-HRP conjugate, the detection range of DON showed 3-3,000 ng/ml (ppb). The monoclonal antibody showed some cross-reactivities against DON analogues such as 15 acetyl-DON (110%), nivalenol (5.0%), 3 acetyl-DON (1.7%), fusarenon-x (0.72%), and T-2 (0.59%). When the cdELISA was applied to the spiked coms after extracting with 60% methanol and diluting 5- fold with washing buffer, the assay recoveries of DON were 313, 163, 106, and 88.9% (av., 168%) in the levels of 200, 600, 2,000, and 6,000ng/g, respectively. For the quantitation of DON in coms, 30 samples kept under two different storage conditions of cold and room temperature were assayed by cdELISA. The mean detection concentrations were 595 (detection range, 0-2,750) and 2,448 (detection range, 0-4,500) ppb, respectively.

Keywords

Deoxynivalenol;Monoclonal antibody;ELISA;Corn

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