High-level production of N-terminal pro-brain natriuretic peptide, as a calibrant of heart failure diagnosis, in Escherichia coli

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dc.contributor.authorKim, Young Suko
dc.contributor.authorKarisa, Nadiako
dc.contributor.authorJeon, Woo Youngko
dc.contributor.authorLee, Hongweonko
dc.contributor.authorKim, Yeu-chunko
dc.contributor.authorAhn, Jungohko
dc.date.accessioned2019-06-19T01:30:07Z-
dc.date.available2019-06-19T01:30:07Z-
dc.date.created2019-06-18-
dc.date.created2019-06-18-
dc.date.issued2019-06-
dc.identifier.citationAPPLIED MICROBIOLOGY AND BIOTECHNOLOGY, v.103, no.12, pp.4779 - 4788-
dc.identifier.issn0175-7598-
dc.identifier.urihttp://hdl.handle.net/10203/262736-
dc.description.abstractHeart failure (HF) is a coronary disease that affects people worldwide and has a high mortality rate. N-terminal pro-brain natriuretic peptide (NT-proBNP) has been proven to be a useful and accurate biomarker for diagnosing systolic HF. Here, we report a strategy for the high-level production of recombinant (r)NT-proBNP in Escherichia coli. An Fh8 tag with six histidines was fused to the N terminus of NT-proBNP along with the recognition site of tobacco etch virus (TEV) protease; the 6HFh8-NT-proBNP fusion peptide was expressed in flask cultures of E. coli in almost completely soluble form. The peptide was purified by HisTrap affinity chromatography, and the N-terminal tag was cleaved by TEV protease. After a second round of HisTrap affinity chromatography to remove the TEV protease and N-terminal tag, rNT-proBNP was isolated with high purity (98%) by carboxymethyl cation exchange chromatography. The final yield of purified rNT-proBNP (97.5mg/l of bacterial culture; 3.25mg/g of wet cell) was 55-fold higher than that reported in previous studies (0.5-1.75mg/l of bacterial culture). Furthermore, the high cell density E. coli fed-batch culture enabled high-level production of rNT-proBNP in the order of grams per liter. The purified rNT-proBNP was detected by enzyme-linked immunosorbent assay and chemiluminescence enzyme immunoassay using commercial monoclonal antibodies recognizing different epitopes, showing a linear dose-response relationship in the range of tested concentrations (slope=3.58 and r(2)=0.995). These results demonstrate the efficiency of our process for mass producing (gram-to-liter level) rNT-proBNP with acceptable analytical performance.-
dc.languageEnglish-
dc.publisherSPRINGER-
dc.titleHigh-level production of N-terminal pro-brain natriuretic peptide, as a calibrant of heart failure diagnosis, in Escherichia coli-
dc.typeArticle-
dc.identifier.wosid000469192100010-
dc.identifier.scopusid2-s2.0-85065221657-
dc.type.rimsART-
dc.citation.volume103-
dc.citation.issue12-
dc.citation.beginningpage4779-
dc.citation.endingpage4788-
dc.citation.publicationnameAPPLIED MICROBIOLOGY AND BIOTECHNOLOGY-
dc.identifier.doi10.1007/s00253-019-09826-8-
dc.contributor.localauthorKim, Yeu-chun-
dc.contributor.nonIdAuthorKarisa, Nadia-
dc.contributor.nonIdAuthorJeon, Woo Young-
dc.contributor.nonIdAuthorLee, Hongweon-
dc.contributor.nonIdAuthorAhn, Jungoh-
dc.description.isOpenAccessN-
dc.type.journalArticleArticle-
dc.subject.keywordAuthorNT-proBNP-
dc.subject.keywordAuthorBiomarker-
dc.subject.keywordAuthorDiagnosis-
dc.subject.keywordAuthorHeart failure-
dc.subject.keywordAuthorEscherichia coli-
dc.subject.keywordPlusA-TYPE-
dc.subject.keywordPlusBNP-
dc.subject.keywordPlusFRAGMENTS-
dc.subject.keywordPlusPROTEINS-
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