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Preparation of Anti-Fumonisin B_1 Single Chain Antibody Fragment(scFv)and Establishment of Detection System for Fumonisin B_1

【摘要】:正Fumonisin is a kind of mycotoxin produced by fusarium verticillioides and fusarium moniliform sheld,mainly in corn and corn-based products.Although several other fumonisin analogues have been characterized,such as FA_2,FA_2,and FB_2.Fumonisin B_1(FB_1) remains the most abundant in naturally contaminated corn-based foods,followed by Fumonisin B_2(FB_2).So far,a lot of studies have been reported using anti-fumonisin B_1 monoclonal antibody to detect FB_1 in crops,and have gained achievements in some extent.ScFv also can be used to achieve the same or better effect,and it can overcome some inherent shortcomings of monoclonal antibodies.In this paper,the hapten fumonisin was coupled to a protein carrier BSA(Bovine Serum Albumin),and the conjugate was used for immunization before extracting mRNA from spleen cells of the immunized mice.Then the gene coding a single variable chain antibody(scFv) against the highly toxic mycotoxin fumonisin B_1 has been assembled and amplified successfully now.The result indicated that the immunological effect was comparatively well,which suggested that the pimary product of scFv experssion could get relatively high titer.After cloning and experssion,the scFv obtained would be applied to construct naive scFv library before three panning rounds.And the antibody after panning can potentially be used for developing a rapid and affordable immunoassay for detection of food contamination.

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