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发明公开
CN103898615A 一种黄曲霉毒素纳米抗体基因库、构建方法、用途及黄曲霉毒素M1纳米抗体2014AFM-G2 有权

一种黄曲霉毒素纳米抗体基因库、构建方法、用途及黄曲霉毒素M1纳米抗体2014AFM-G2
摘要：
本发明涉及黄曲霉毒素纳米抗体基因库、构建方法、用途及黄曲霉毒素M1纳米抗体2014AFM-G2。所述黄曲霉毒素纳米抗体基因库是通过提取免疫黄曲霉毒素M1完全抗原后的羊驼血液中的RNA，并采用反转录PCR的方法特异性扩增羊驼重链抗体可变区基因得到黄曲霉毒素纳米抗体VHH基因，然后与pCANTAB5E（his）载体连接后转化得到。本发明筛选得到的黄曲霉毒素M1纳米抗体2014AFM-G2具有耐有机试剂、耐高温、耐酸碱等特点，稳定性好；对黄曲霉毒素M1的50%抑制浓度IC50为0.208ng/mL；与黄曲霉毒素B1，B2，G1，G2交叉反应率分别为9.43%、5.93%、4.87%和6.17%。
摘要（英）：

The invention relates to an aflatoxin nanometer antibody gene bank, a construction method thereof and an application thereof as well as an aflatoxin M1 nanometer antibody 2014AFM-G2. The aflatoxin nanometer antibody gene bank is obtained by adopting the following steps: extracting RNA in alpaca blood after immunizing complete antigen of aflatoxin M1, specifically amplifying genes in a heavy-chain antibody variable region of alpaca by using a reverse transcription PCR (Polymerase Chain Reaction) method to obtain aflatoxin nanometer antibody VHH (variable domain of the heavy-chain of heavy-chain antibody) genes, then connecting the aflatoxin nanometer antibody VHH genes and pCANTAB5E(his) carriers, and transforming. The aflatoxin M1 nanometer antibody 2014AFM-G2 obtained by screening has good stability and characteristics of organic reagent resistance, high temperature resistance, acid-base resistance and the like. The 50% inhibition concentration IC50 of the antibody to the aflatoxin M1 is 0.208ng/mL; the cross reaction rates between the antibody and aflatoxin B1, between the antibody and aflatoxin B2, between the antibody and the aflatoxin G1 and between the antibody and the aflatoxin G2 are respectively 9.43%, 5.93%, 4.87% and 6.17%.

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