A rapid and cost-effective method of producing recombinant proBNP and NT-proBNP variants in Escherichia coli for immunoassay of heart failure
Biotechnology Letters, ISSN: 0141-5492, Vol: 36, Issue: 1, Page: 133-140
2014
- 4Citations
- 11Captures
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Metrics Details
- Citations4
- Citation Indexes4
- CrossRef2
- Captures11
- Readers11
- 11
Article Description
The measurements of plasma natriuretic peptides (NT-proBNP, proBNP and BNP) are used to diagnose heart failure but these are expensive to produce. We describe a rapid, cheap and facile production of proteins for immunoassays of heart failure. DNA encoding N-terminally His-tagged NT-proBNP and proBNP were cloned into the pJexpress404 vector. ProBNP and NT-proBNP peptides were expressed in Escherichia coli, purified and refolded in vitro. The analytical performance of these peptides were comparable with commercial analytes (NT-proBNP EC for the recombinant is 2.6 ng/ml and for the commercial material is 5.3 ng/ml) and the EC for recombinant and commercial proBNP, are 3.6 and 5.7 ng/ml respectively). Total yield of purified refolded NT-proBNP peptide was 1.75 mg/l and proBNP was 0.088 mg/l. This approach may also be useful in expressing other protein analytes for immunoassay applications. Purpose of work: To develop a cost effective protein expression method in E. coli to obtain high yields of NT-proBNP (1.75 mg/l) and proBNP (0.088 mg/l) peptides for immunoassay use. © 2013 Springer Science+Business Media Dordrecht.
Bibliographic Details
http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=84891652144&origin=inward; http://dx.doi.org/10.1007/s10529-013-1341-0; http://www.ncbi.nlm.nih.gov/pubmed/24101238; http://link.springer.com/10.1007/s10529-013-1341-0; https://dx.doi.org/10.1007/s10529-013-1341-0; https://link.springer.com/article/10.1007/s10529-013-1341-0
Springer Science and Business Media LLC
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