A quantitative ELISA for monitoring the secretion of ZZ-Fusion proteins using SpA domain as immunodetection reporter system

摘要

A sandwich-type enzyme-linkd immunosorbent assay (ELISA) was estabilshed for monitoring the secretion of ZZ-fusion proteins.Two antibodies,a monoclonal mouse anti-human proinsulin and a rabbit antibovine IgG (strongly binding to theZZ-domain),were used to quatify thesecretion of recombinant human ZZ-proinsulin to te growth medium of Escherichia coli cultures.The method here reported conjugates the advantages of sandwich-type ELISA assays,namely,high sensitivity,specificity,and hroughput,with the possibility of quantifying small protein moelcules (e.g.,peptides).A further advantage of gene fusion techniques integrating both downstream processing and product detection and quantitation is highlighted.The method is capable of detecting levels of 0.05 ng of ZZ-proinsulin.