使用三种单克隆抗体混合物提高人α1干扰素酶免疫测定的灵敏度。

Increased sensitivity of an enzyme immunoassay for human interferon alpha 1 using a mixture of three monoclonal antibodies.

作者信息

Karayianni-Vasconcelos G, Mucha V, Kontsek P

机构信息

Institute of Virology, Slovak Academy of Sciences, Bratislava.

出版信息

J Immunoassay. 1994 Aug;15(3):251-61. doi: 10.1080/15321819408009576.

DOI:10.1080/15321819408009576

PMID:7929852

Abstract

A sensitive sandwich ELISA for the quantification of human interferon (IFN)-alpha 1 was designed. The assay employed IFN-alpha 1- specific polyclonal antibody for coating and monoclonal antibodies (mAbs) to IFN-alpha 1 as a second antibody. A major increase in sensitivity of the assay could be achieved, when polyclonal antibody was combined with a mixture of three mAbs binding to different regions of IFN-alpha 1, compared to the combination of a polyclonal antibody with a single mAb. The sensitivity of the established ELISA was close to that of an antiviral IFN-bioassay. The ELISA did not cross-react with IFN-alpha 2, beta, tau or omega. The immunoassay allowed to estimate the content of IFN-alpha 1 in leukocyte IFN-alpha to about 25-50% or to 2-6% in Namalwa IFN-alpha, respectively.

摘要

设计了一种用于定量人α1干扰素（IFN-α1）的灵敏夹心酶联免疫吸附测定（ELISA）法。该检测采用针对IFN-α1的特异性多克隆抗体进行包被，并使用抗IFN-α1单克隆抗体（mAb）作为第二抗体。与多克隆抗体与单个mAb组合相比，当多克隆抗体与结合到IFN-α1不同区域的三种mAb混合物联合使用时，该检测的灵敏度可实现大幅提高。所建立的ELISA的灵敏度与抗病毒IFN生物测定法相近。该ELISA与IFN-α2、β、τ或ω无交叉反应。该免疫测定法能够分别估计白细胞IFN-α中IFN-α1的含量约为25%-50%，在Namalwa IFN-α中为2%-6%。