α-乳白蛋白单克隆抗体的制备及斑点印迹定性检测方法的建立  

Development of monoclonal antibodies against α-lactalbumin and establishment of qualitative detection for α- lactalbumin with dot blotting assay

作　　者：龙彩云[1,2] 郑义成[1,2] 程芬芬[1,2] 杨安树[1,2] 陈红兵[1,2] 

机构地区：[1]南昌大学食品科学与技术国家重点实验室,江西南昌330047 [2]南昌大学中德联合研究院,江西南昌330047

出　　处：《食品工业科技》2014年第6期185-188,共4页Science and Technology of Food Industry

基　　金：国家高技术研究发展计划(863计划)(2013AA102205);国家科技支撑计划项目(2011BAK10B03);江西省青年科学家培养对象计划(20122BCB23006);江西省自然科学基金资助项目(20122BAB204001)

摘　　要：以α-乳白蛋白为抗原免疫Balb/c小鼠,采用淋巴细胞杂交瘤技术制备抗α-乳白蛋白单克隆抗体;采用碳二亚胺法,并通过工艺优化,将单抗与量子点高效偶联;在此基础上,建立了基于斑点印迹技术定性检测乳制品中过敏原α-乳白蛋白的方法。结果表明:采用杂交瘤技术制备的单克隆抗体特异性好,与牛奶中其他蛋白无交叉反应;当量子点、1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(EDC)、N-羟基丁二酰亚胺(NHS)、单抗摩尔比为1∶10∶6∶4时,量子点与单抗有较好的偶联效果;以量子点标记单抗建立的斑点印迹方法可直观、快速的定性分析乳制品中过敏原α-乳白蛋白。因此,该方法具有一定的实用价值。The monoclonal antibodies（mAbs） against bovine α-lactalbumin（α-la） were prepared by immunning Balb/c mice and the lymphocyte hybridoma technique. The mAbs were covalently conjugated with the CdSe/ ZnS quantum dots （QDs） by the carbodiimide crosslinking method,and then the coupling conditions were optimized. Furthmore,a dot blotting method based on QD-mAb conjugates was established to qualitatively detect α-La in different commercial dairy products. The results showed that the prepared mAbs had high specificity and no cross-reaction. A good coupling effect was confirmed when the molar ratio of QD,N-（3- Dimethylaminopropyl ） - N ＇ -ethylcarbodiimide hydrochloride （EDC）, N-Hydroxysuccinimide （NHS）, monoclonal antibody was 1：10：6：4. The developed method based on QD-mAb conjugates could be used to visual,rapid qualitative analyze α-La in dairy products. Accordingly,the method possesses certain practical value.

关 键 词：Α-乳白蛋白 单克隆抗体 量子点 斑点印迹 定性检测 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]