Highly Sensitive Enzyme Immunoassay of Human Interferon-β1

Abstract

A highly sensitive sandwich enzyme immunoassay for human interferon-β 1(HuIFN-β 1) was developed. HuIFN-β 1-containing samples and horseradish peroxidase (HRP)-labeled mouse anti-HuIFN-β1 monoclonal antibody (Fab') were incubated overnight at 2–10°C in the wells of a 96-well microtiter plate, onto which affinity-purified rabbit anti-HuIFN-β 1 polyclonal antibody was coated. The EIA was able to detect 0.5 IU/m1 of HuIFN-β1, thus showing higher sensitivity than bioassay. The values obtained by the EIA closely paralleled those obtained by bioassay in the concentration which bioassay can detect. In order to detect the concentration below 0.5 IU/ml of HuIFN-β 1, the avidin/biotin-amplified EIA was also developed. The use of biotinylated mouse anti-HuIFN-β 1 monoclonal antibody (F(ab')2) and HRP-avidin in the EIA made it possible to detect 0.1 IU/ml of HuIFN-β1. These EIAs were applied for the studies such as process control of HuIFN-β 1 production, pharmacokinetics of HuIFN-β 1, and determination of serum level of HuIFN-β1 in healthy subjects.