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核农学报  2015 

玉米赤霉醇特异性单克隆抗体及其胶体金检测试纸的研制

DOI: 10.11869/j.issn.100-8551.2015.06.1101, PP. 1101-1107

Keywords: 玉米赤霉醇,单克隆抗体,玉米赤霉醇-16-羧丙基醚,玉米赤霉烯酮,胶体金检测试纸

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Abstract:

为了研究国内制备的玉米赤霉醇单克隆抗体,与其同系物玉米赤霉烯酮交叉反应严重,实际检测存在假阳性的问题,本试验开展了玉米赤霉醇特异性识别单克隆抗体及其胶体金检测试纸的研制工作。试验采用玉米赤霉醇-16-羧丙基醚与牛血清白蛋白偶联,免疫雌性Balb/c系小鼠;利用单克隆抗体技术完成杂交瘤融合,并用非竞争及竞争ELISA法,对阳性克隆进行鉴定。杂交瘤诱生腹水后,经亲和纯化,用于15nm胶体金标记,制作玉米赤霉醇胶体金检测试纸。融合细胞经筛选,最终对3株杂交瘤细胞建株,结果均为IgG1型。其中Mab-1A8诱生腹水效价高达1:1.28×107,对玉米赤霉醇抑制中浓度(I50)为10.25ng·mL-1,检测灵敏度(I10)为1.96ng·mL-1,线性检测范围在2.96~35.46ng·mL-1,对同系物玉米赤霉烯酮的交叉反应率小于0.1%。试验研制的玉米赤霉醇胶体金试纸,检测阈值为2.5μg·mL-1,检测时间为10min。本研究制备了一种新型玉米赤霉醇特异性单克隆抗体,并研制了以单抗为基础的玉米赤霉醇胶体金快速检测试纸,为玉米赤霉醇残留快速检测提供了灵敏、便捷的新方法。

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