抗桔青霉素单克隆抗体的研制与鉴定

目的制备抗桔青霉素的单克隆抗体。方法采用活性酯法、甲醛加成法和羰基二咪唑法制备了4种桔青霉素与载体蛋白的偶联物(A、B、C和D),免疫小鼠制备抗桔青霉素单克隆抗体。结果免疫原性鉴定证明偶联物C可刺激BALB/c小鼠产生抗桔青霉素的抗体,细胞融合后筛选到一株抗桔青霉素的单克隆抗体,单抗与赭曲霉毒素A、黄曲霉毒素B1和展青霉素等毒素交叉反应低于0.01%,在此基础上建立了竞争ELISA检测方法,线性范围为20~10,00ng/ml,检测下限为10ng/ml。在小麦样品中的加标回收率为95%~112%,变异系数为9.1%~18.6%。结论本研究成功筛选到抗桔青霉素单克隆抗体。

Preparation and identification of a monoclonal antibody against citrinin

OBJECTIVE: To prepare the monoclonal antibody (McAb) against citrinin. METHODS: Four citrinin-protein conjugates (A, B, C and D) were synthesized by the active ester, Munnich Reaction and 1,1-carbonyldiimidazole (CDI) method, respectively. BALB/c mice were immunized with the conjugates for producing the McAb against citrinin. RESULTS: The antibodies against citrinin have been produced in serum of immunized BALB/c mice with the conjugate C. A McAb against citrinin was acquired according to the traditional procedure. Cross reactivity of the McAb was less than 0.01% with ochratoxin A, patulin, aflatoxin B1, deoxynivalenol and zearalenone, respectively. A competitive enzyme-linked immunosorbent assay was developed. The linear range of ELISA was between 20 ng/ml and 1 000 ng/ml, the detectable limit was 10 ng/ml. Recovery of citrinin from wheat spiked with citrinin was from 95% to 112%, the coefficient of variation was 12.2% to 20.4%. CONCLUSION: The McAb against citrinin was acquired successfully.