A rapid and cost-effective method of producing recombinant proBNP and NT-proBNP variants in Escherichia coli for immunoassay of heart failure
Data(s) |
2014
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Resumo |
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. |
Identificador | |
Publicador |
Springer |
Relação |
DOI:10.1007/s10529-013-1341-0 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. |
Fonte |
School of Biomedical Sciences; Faculty of Health |
Palavras-Chave | #AlphaLISA #Escherichia coli #Heart failure #Immunoassay #Natriuretic #peptides #Plasma peptides #natriuretic peptide #terminal fragments #a-type #guidelines #society #saliva #corin #assay |
Tipo |
Journal Article |