DC Field | Value | Language |
---|---|---|
dc.contributor.author | Kim, Young Su | ko |
dc.contributor.author | Karisa, Nadia | ko |
dc.contributor.author | Jeon, Woo Young | ko |
dc.contributor.author | Lee, Hongweon | ko |
dc.contributor.author | Kim, Yeu-chun | ko |
dc.contributor.author | Ahn, Jungoh | ko |
dc.date.accessioned | 2019-06-19T01:30:07Z | - |
dc.date.available | 2019-06-19T01:30:07Z | - |
dc.date.created | 2019-06-18 | - |
dc.date.created | 2019-06-18 | - |
dc.date.issued | 2019-06 | - |
dc.identifier.citation | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, v.103, no.12, pp.4779 - 4788 | - |
dc.identifier.issn | 0175-7598 | - |
dc.identifier.uri | http://hdl.handle.net/10203/262736 | - |
dc.description.abstract | Heart 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.language | English | - |
dc.publisher | SPRINGER | - |
dc.title | High-level production of N-terminal pro-brain natriuretic peptide, as a calibrant of heart failure diagnosis, in Escherichia coli | - |
dc.type | Article | - |
dc.identifier.wosid | 000469192100010 | - |
dc.identifier.scopusid | 2-s2.0-85065221657 | - |
dc.type.rims | ART | - |
dc.citation.volume | 103 | - |
dc.citation.issue | 12 | - |
dc.citation.beginningpage | 4779 | - |
dc.citation.endingpage | 4788 | - |
dc.citation.publicationname | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY | - |
dc.identifier.doi | 10.1007/s00253-019-09826-8 | - |
dc.contributor.localauthor | Kim, Yeu-chun | - |
dc.contributor.nonIdAuthor | Karisa, Nadia | - |
dc.contributor.nonIdAuthor | Jeon, Woo Young | - |
dc.contributor.nonIdAuthor | Lee, Hongweon | - |
dc.contributor.nonIdAuthor | Ahn, Jungoh | - |
dc.description.isOpenAccess | N | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordAuthor | NT-proBNP | - |
dc.subject.keywordAuthor | Biomarker | - |
dc.subject.keywordAuthor | Diagnosis | - |
dc.subject.keywordAuthor | Heart failure | - |
dc.subject.keywordAuthor | Escherichia coli | - |
dc.subject.keywordPlus | A-TYPE | - |
dc.subject.keywordPlus | BNP | - |
dc.subject.keywordPlus | FRAGMENTS | - |
dc.subject.keywordPlus | PROTEINS | - |
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