重组人γ干扰素酶联免疫吸附测定法（ELISA）的开发与验证

Development and validation of an enzyme-linked immunosorbent assay (ELISA) for recombinant human gamma interferon.

作者信息

Bouyón R, Santana H, Pérez E M, Hernández N, Furrazola G, Abrahantes M C

机构信息

Center for Genetic Engineering and Biotechnology, Havana, Cuba.

出版信息

J Immunoassay Immunochem. 2003;24(1):1-10. doi: 10.1081/IAS-120018465.

DOI:10.1081/IAS-120018465

PMID:12680603

Abstract

A one-site ELISA for the quantification of recombinant human gamma interferon (rh-IFN-gamma) was developed and validated. A single monoclonal antibody (Mab) was used as a "catching" antibody and as a horseradish peroxidase (HRP)-labeled conjugate. Detection limit and quantification limit of this assay were estimated to be 1.26 and 15 ng/mL, respectively, and the coefficient of variation was below 15%. The ELISA was specific for rh-IFN-gamma, showing no cross reactivity to other related molecules in the range of the concentrations studied. The results correlated well with those obtained by a bioassay method. By using this assay, it was demonstrated that 0.01-1% (v/v) Tween 80 protected rh-IFN-gamma during freezing and thawing.

摘要

开发并验证了一种用于定量重组人γ干扰素（rh-IFN-γ）的单位点酶联免疫吸附测定（ELISA）。使用一种单克隆抗体（Mab）作为“捕获”抗体以及辣根过氧化物酶（HRP）标记的缀合物。该测定法的检测限和定量限分别估计为1.26和15 ng/mL，变异系数低于15%。该ELISA对rh-IFN-γ具有特异性，在所研究的浓度范围内对其他相关分子无交叉反应。结果与通过生物测定法获得的结果高度相关。通过使用该测定法，证明0.01 - 1%（v/v）的吐温80在冻融过程中对rh-IFN-γ具有保护作用。