Production of Monoclonal Antibody and Development of ELISA and Gold Nanoparticle Immunochromatographic Strip Assay for Aflatoxin B1

• Main Authors: Jiun-wei Liu, 劉俊緯
• Other Authors: Feng-Yih Yu
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/92969354657536662841

碩士 === 中山醫學大學 === 生物醫學科學學系碩士班 === 101 === Mycotoxins are secondary metabolites produced by fungi. Foods or feeds which are contaminated by mycotoxins will harm the health of people or animals by ingestion. Aflatoxins, including AFB1、B2、G1、G2, were the most commonly occurred mycotoxins in cereal grain. According to previous research, AFB1 is the most toxic one which cause acute hepatitis or hepatic cancer by its hepatotoxicity. International Agency for Research on Cancer (IARC) has classified AFB1 as group 1, as a human carcinogen. Therefore, the U.S. Food and Drug Administration (FDA) enacted a regulatory limit of total aflatoxins in foods and feeds as 20 ppb. The maximum levels of total aflatoxins in Taiwan is 15 ppb. Moreover, AFB1 should below 2 ppb in the European Union. For correct and rapid detecting AFB1 in food and feeds, we produced AFB1-specific monoclonal antibody and applied it on a competitive direct enzyme-linked immunosorbent assay (cdELISA) and a gold nanoparticle based rapid immunochromatographic strip. The concentration causing 50% inhibition of binding AFB1-CMO-HRP to AFB1 antibodies (IC50) in cdELISA were found to be 0.0515 ng/mL in 3F6G11 and 0.0451 ng/mL in 9C7C11H10. The detection limit of immunochromatographic strip for AFB1 were 2 ng/mL in 3F6G11 and 1 ng/mL in 9C7C11H10, respectively. Results of analyzing food and feeds based on 3F6G11 monoclonal antibody showed a good agreement in cdELISA and strip. These data suggested that we successfully developed two kinds of rapid immunoassay by using AFB1 monoclonal antibody 3F6G11, cdELISA and strip assay, which could simply and rapidly detect AFB1 contamination in foods and feeds.