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

Abstract:

:A sandwich-type enzyme-linked immunosorbent assay (ELISA) was established for monitoring the secretion of ZZ-fusion proteins. Two antibodies, a monoclonal mouse anti-human proinsulin and a rabbit anti-bovine IgG (strongly binding to the ZZ-domain), were used to quantify the secretion of recombinant human ZZ-proinsulin to the growth medium of Escherichia coli cultures. The method here reported conjugates the advantages of sandwich-type ELISA assays, namely, high sensitivity, specificity, and throughput, with the possibility of quantifying small protein molecules (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.

journal_name

Mol Biotechnol

journal_title

Molecular biotechnology

authors

Mergulhão FJ,Monteiro GA,Cabral JM,Taipa MA

doi

10.1385/MB:19:3:239

keywords:

subject

Has Abstract

pub_date

2001-11-01 00:00:00

pages

239-44

issue

3

eissn

1073-6085

issn

1559-0305

pii

MB:19:3:239

journal_volume

19

pub_type

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