酶联免疫吸附测定中封闭液的作用与选择
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摘要
酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)的基础是抗原或抗体的固相化和抗原或抗体的酶标记。测定时,受检标本(测定其中的抗体或抗原)需先与固相载体表面的抗原或抗体起反应,结合在固相载体表面(包被)。由于抗原或抗体包被时的浓度较低,故还需用大量不相关的蛋白充填固相载体表面空隙(封闭),使之避免在ELISA其后步骤中吸附干扰物质而影响测定。本文概要介绍ELISA中各种常用封闭液的作用与选择。
Enzyme-linked immunosorbent assay(ELISA) is based on the immobilization of antigen or antibody and the enzymatic labeling of antigen or antibody. At the time of determination, the specimen to be examined(antibody or antigen therein) needs to first react with the antigen or antibody on the surface of the solid support and then bind to the surface of the solid support(coating). Due to the low concentrations of the coated antigen or antibody, a large amount of irrelevant proteins are also needed to fill the spacing on the solid support surface(closed), thus to avoid adsorption of interfering substances that may affect the determination in the subsequent steps of the ELISA. The role and selection of various commonly used blocking solutions in ELISA are outlined in this article.
Enzyme-linked immunosorbent assay(ELISA) is based on the immobilization of antigen or antibody and the enzymatic labeling of antigen or antibody. At the time of determination, the specimen to be examined(antibody or antigen therein) needs to first react with the antigen or antibody on the surface of the solid support and then bind to the surface of the solid support(coating). Due to the low concentrations of the coated antigen or antibody, a large amount of irrelevant proteins are also needed to fill the spacing on the solid support surface(closed), thus to avoid adsorption of interfering substances that may affect the determination in the subsequent steps of the ELISA. The role and selection of various commonly used blocking solutions in ELISA are outlined in this article.
引文
[1]焦奎,张书圣.酶联免疫分析技术及应用[M].北京:化学工业出版社,2004:96-107.
[2]Gibbs J.Effective blocking procedures.ELISA technical bulletin No.3[EB/OL].[2018-01-17].http://www.doc88.com/p-7864239831977.html.
[3]李秀玲,倪道明.人抗动物抗体对免疫学测定的干扰作用[J].国外医学:预防、诊断、治疗用生物制品分册,2001,24(6):266-270.
[4]刘苏燕,董邦全,周雪慧,等.吐温20、BSA和8种血清在ELISA中抑制作用的比较[J].天津医药,1991,19(6):373-374.
[2]Gibbs J.Effective blocking procedures.ELISA technical bulletin No.3[EB/OL].[2018-01-17].http://www.doc88.com/p-7864239831977.html.
[3]李秀玲,倪道明.人抗动物抗体对免疫学测定的干扰作用[J].国外医学:预防、诊断、治疗用生物制品分册,2001,24(6):266-270.
[4]刘苏燕,董邦全,周雪慧,等.吐温20、BSA和8种血清在ELISA中抑制作用的比较[J].天津医药,1991,19(6):373-374.