Quantitative detection of fusion protein rIFN-ß-HSA by a sandwich ELISA

Abstract

A method that allowed detection and quantification of recombinant fusion protein rIFN-ß-HSA in complex mixtures and fermentation media was described. The method was based on a double antibody sandwich ELISA, which was developed by using an anti-IFN-ß monoclonal antibody as capture antibody and an HRP-labeled anti- human serum albumin (HSA) monoclonal antibody as detection antibody. The practical working range was estimated to be 21.01 ng/ml to 672.50 ng/ml and the limit of detection was 13.88 ng/ml. Recoveries ranged from 91.9 to 110.4%, while the intra and inter-assay precisions were <4.86 and <8.08%, respectively.

Key words: ELISA, rIFN-ß, human serum albumin (HSA), Pichia pastoris, fermentation.