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, & 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), pp. 133-140.
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. 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.
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|Item Type:||Journal Article|
|Additional Information:||<Go to ISI>://WOS:000329228800018|
|Keywords:||AlphaLISA, Escherichia coli, Heart failure, Immunoassay, Natriuretic, peptides, Plasma peptides, natriuretic peptide, terminal fragments, a-type, guidelines, society, saliva, corin, assay|
|Divisions:||Current > Schools > School of Biomedical Sciences
Current > QUT Faculties and Divisions > Faculty of Health
|Deposited On:||23 Oct 2014 04:35|
|Last Modified:||23 Oct 2014 22:00|
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