高效毛细管电泳测定纳米抗体亲和力  

Determination of Affinity of Nano-antibody by High Performance Capillary Electrophoresis

作　　者：张腾飞 韩雪容 李飞 于建立 宋海鹏 ZHANG Teng-fei;HAN Xue-rong;LI Fei;YU Jian-li;SONG Hai-peng(School of Life Science and Technology,Changchun University of Science and Technology,Changchun 130022;Shenzhen Innova Nanobodi Co.,Ltd,Shenzhen 518000;CAS Key Laboratory of Biobased Materials,Qingdao Institute of Bioenergy and Bioprocess Technology,Chinese Academy of Sciences,Qingdao 266101)

机构地区：[1]长春理工大学生命科学技术学院,长春130022 [2]深圳市国创纳米抗体技术有限公司,深圳518000 [3]中国科学院青岛生物能源与过程研究所,青岛266101

出　　处：《长春理工大学学报（自然科学版）》2019年第5期138-142,共5页Journal of Changchun University of Science and Technology(Natural Science Edition)

基　　金：吉林省重点科技攻关项目（20170204044GX)

摘　　要：通过毛细管电泳检测技术,应用到检测纳米抗体亲和力上,找到新的检测纳米抗体亲和力测定的方法。利用毛细管电泳在分析检测上分离速度快、实验时间短、样品和溶剂用量少的优势,在最佳浓度、pH、电压条件下来测定纳米抗体和抗原的结合常数。通过实验不断优化毛细管电泳浓度、pH和电压的条件,使毛细管电泳测定纳米抗体达到最佳测试状态,通过最优化条件实验表明,在电泳浓度为50 mM、pH=7和电压为20 kV的条件时为最佳测试实验条件,通过不同浓度抗体和定值抗原的结合,绘制标准曲线并计算出测定常数,求得纳米抗体抗原的结合常数,并用现在常用的Biacore检测方法对实验结果进行验证。结果表明,优化条件后毛细管电泳在测定纳米抗体亲和力方面与Biacore测定结果相近,为以后纳米抗体亲和力测定提供一种新方法。In this paper,a new method was found to detect the affinity of nano-antibody by capillary electrophoresis.Capillary electrophoresis was used to determine the binding constants of nano-antibodies and antigens under the optimum concentration and PH,voltage,such as high separation speed,short experimental time and less amount of sample and solvent.The conditions of capillary electrophoretic concentration,pH and voltage were optimized to make the determination of nano-antibody by capillary electrophoresis to be the best test state.The optimum conditions showed that the electrophoresis concentration was 50 mM pH=(7)and the voltage was 20 kV.According to the combination of different concentration of antibody and fixed value antigen,the standard curve was drawn and the determination constant was calculated;and the binding constant of nano-antibody antigen was obtained.The experimental results were verified by the current Biacore detection method.The results showed that the results of capillary electrophoresis were similar to those of Biacore in the determination of the affinity of nano-antibodies,which provided a new method for the determination of affinity of nano-antibodies in the future.

关 键 词：毛细管电泳 检测 纳米抗体 亲和力 

分 类 号：Q816[生物学—生物工程]