A rapid and cost-effective method of producing recombinant proBNP and NT-proBNP variants in Escherichia coli for immunoassay of heart failure

Soleh, Muhammad Tarmizi, Foo, Jared Yong Yang, Bailey, Ulla-Maja, Tan, Nikki Yi, Wan, Yunxia, Cooper-White, Justin, Schulz, Benjamin Luke and Punyadeera, Chamindie (2014) A rapid and cost-effective method of producing recombinant proBNP and NT-proBNP variants in Escherichia coli for immunoassay of heart failure. Biotechnology Letters, 36 1: 133-140. doi:10.1007/s10529-013-1341-0


Author Soleh, Muhammad Tarmizi
Foo, Jared Yong Yang
Bailey, Ulla-Maja
Tan, Nikki Yi
Wan, Yunxia
Cooper-White, Justin
Schulz, Benjamin Luke
Punyadeera, Chamindie
Title A rapid and cost-effective method of producing recombinant proBNP and NT-proBNP variants in Escherichia coli for immunoassay of heart failure
Formatted title
A rapid and cost-effective method of producing recombinant proBNP and NT-proBNP variants in Escherichia coli for immunoassay of heart failure
Journal name Biotechnology Letters   Check publisher's open access policy
ISSN 0141-5492
1573-6776
Publication date 2014
Year available 2013
Sub-type Article (original research)
DOI 10.1007/s10529-013-1341-0
Volume 36
Issue 1
Start page 133
End page 140
Total pages 8
Place of publication Dordrecht, Netherlands
Publisher Springer
Collection year 2014
Language eng
Subject 1305 Biotechnology
Formatted abstract
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 EC50 for the recombinant is 2.6 ng/ml and for the commercial material is 5.3 ng/ml) and the EC50 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. 
Keyword AlphaLISA
Escherichia coli
Heart failure
Immunoassay
Natriuretic peptides
Plasma peptides
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Published online: 8 October 2013.

 
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