抗鸡蛋清提取物单克隆抗体的制备与鉴定

食品科学 ›› 2008, Vol. 29 ›› Issue (10): 372-374.

抗鸡蛋清提取物单克隆抗体的制备与鉴定

许静, 傅颖媛, 刘志刚, 李佳娜

南方医科大学华瑞医院；南昌大学医学院免疫学教研室；深圳大学生命科学学院变态反应学实验室；

出版日期:2008-10-15 发布日期:2011-12-08

Preparation and Characterization of Monoclonal Antibodies Against Purified Egg White Extract

XU Jing, FU Ying-Yuan, LIU Zhi-Gang, LI Jia-Na

1. Huarui Hospital,Southern Medical University,Guangzhou 510360,China; 2.Teaching and Research Section of Immunology,Medical College,Nanchang University,Nanchang 330006,China; 3.Research Institute of Allergy,College of Life Science,Shenzhen University,Shenzhen 518060,China

Online:2008-10-15 Published:2011-12-08

摘要/Abstract

摘要： 目的:制备和鉴定小鼠抗鸡蛋清提取物单克隆抗体。方法:以纯化后的蛋清提取物为免疫原免疫BALB/c小鼠,取免疫小鼠脾细胞与小鼠骨髓瘤NS-1细胞融合。通过间接ELISA法筛选可特异性分泌抗蛋清提取物McAb的杂交瘤细胞。采用Ig类与亚类鉴定试剂盒鉴定McAb的Ig类与亚类;通过间接ELISA、Western Blotting鉴定McAb的特异性。结果:获得2株可稳定分泌抗蛋清提取物McAb的的杂交瘤细胞,分别命名为5E12和4F1,其分泌的McAb均为IgG1。Western Blotting分析表明,McAb 4F1所识别的靶分子的相对分子质量(Mr)约为44kD。间接ELISA法检测两株McAb5E12、4F1的冻融前后效价分别为1:3000、1:300。结论:成功制备了两株抗蛋清提取物McAb、5E12和4F1,为变应原的提取、纯化及鉴定提供了有效的实验基础。

关键词: 蛋清提取物, 单克隆抗体, 特性鉴定

Abstract: Objective: To prepare and identify monoclonal antibodies (McAbs) against purified egg white extract. Methods: BALB/c mice were immunized with purified extracts of eggs,and hybridoma cell lines were obtained by fusing immunized mouse spleen cells with myeloma NS-1 cells. The specificities of McAbs were characterized by indirect ELISA and Western Blotting. Results: Two hybridoma cells,5E12 and 4F1,which secreted the McAbs (both are IgG1) against extracts of eggs,are obtained. The titers of 5E12 and 4F1 are 1:3000 and 1:30,respectively. Western Blotting analysis showed that 4F1 can recognize a target molecule with relative molecular mass of 45 kD. The tesults of indirect ELISA showed that the titers of two McAbs are 1:3000 and 1:300,respectively. Conclusion: Two McAbs,5E12 and 4F1,against extracts of eggs are prepared successfully,which provides valuable experiment foundation for quickly measuring allergen in foods as well as for isolating,purifying and identifying allergens.

Key words: purified extracts of eggs, monoclonal antibodies, characterization

许静, 傅颖媛, 刘志刚, 李佳娜. 抗鸡蛋清提取物单克隆抗体的制备与鉴定[J]. 食品科学, 2008, 29(10): 372-374.

XU Jing, FU Ying-Yuan, LIU Zhi-Gang, LI Jia-Na. Preparation and Characterization of Monoclonal Antibodies Against Purified Egg White Extract[J]. FOOD SCIENCE, 2008, 29(10): 372-374.

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