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

• Type: Article in International Scientific Journal
• Date: 2001

Authors

Mergulhao, FJM (Author) Other; Monteiro, GA (Author) Other; Cabral, JMS (Author) Other; Taipa, MA (Author) Other

Journal

Title: Molecular Biotechnology

Vol. 19

Pages: 239-244

ISSN: 1073-6085

Publisher: Springer Nature

Indexing

ISI Web of Knowledge - 11 Citations

ISI Web of Science

Scopus - 12 Citations

Scientific classification

FOS: Natural sciences > Biological sciences

CORDIS: Technological sciences > Technology > Biotecnology

Other information

Authenticus ID: P-000-SYK

DOI: 10.1385/mb:19:3:239

Abstract (EN): 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 antibovine 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.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 6