Title |
A rapid and cost-effective method of producing recombinant proBNP and NT-proBNP variants in Escherichia coli for immunoassay of heart failure
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Published in |
Biotechnology Techniques, October 2013
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DOI | 10.1007/s10529-013-1341-0 |
Pubmed ID | |
Authors |
Muhammad Tarmizi Soleh, Jared Yong Yang Foo, Ulla-Maja Bailey, Nikki Yi Tan, Yunxia Wan, Justin Cooper-White, Benjamin Luke Schulz, Chamindie Punyadeera |
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. |
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United States | 1 | 100% |
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Members of the public | 1 | 100% |
Mendeley readers
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Unknown | 11 | 100% |
Demographic breakdown
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Student > Ph. D. Student | 4 | 36% |
Student > Master | 2 | 18% |
Researcher | 1 | 9% |
Unknown | 4 | 36% |
Readers by discipline | Count | As % |
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Biochemistry, Genetics and Molecular Biology | 3 | 27% |
Medicine and Dentistry | 2 | 18% |
Social Sciences | 1 | 9% |
Nursing and Health Professions | 1 | 9% |
Unknown | 4 | 36% |