| Fast quantitation of recombinant plasminogen activator inhibitor type 1 in bacterial lysates by micropellicular reversed-phase liquid chromatography. | |
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MedLine Citation:
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PMID: 10999627 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A rapid reversed-phase HPLC assay is described for quantitating recombinant plasminogen activator inhibitor type 1 (PAI-1) in cultures of Escherichia coli. The assay utilized a short nonporous micropellicular C18 column with an acetonitrile gradient in 0.1% trifluoroacetic acid. The selective resolution of recombinant PAI-1 from major host proteins occurred within 1.3 min. Regeneration of initial chromatography conditions was fast and allowed successive injections every 3 min. The assay's limit of detection for recombinant PAI-1 was 0.5 microg in 5-microl injections of bacterial lysates and the assay was linear to 20 microg. The assay's apparent recovery was between 94 and 108% throughout the quantitative range. In a direct comparison to gel electrophoresis the reversed-phase assay proved superior in monitoring recombinant PAI-1 titers in cultures of E. coli. |
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Authors:
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D O O'Keefe; G J Duke |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Journal of chromatography. A Volume: 891 ISSN: 0021-9673 ISO Abbreviation: J Chromatogr A Publication Date: 2000 Sep |
Date Detail:
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Created Date: 2001-01-02 Completed Date: 2001-01-11 Revised Date: 2009-01-15 |
Medline Journal Info:
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Nlm Unique ID: 9318488 Medline TA: J Chromatogr A Country: NETHERLANDS |
Other Details:
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Languages: eng Pagination: 85-92 Citation Subset: IM |
Affiliation:
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Department of Macromolecular Structure and Biopharmaceuticals, Bristol-Myers Squibb, Princeton, NJ 08543, USA. okeefed@bms.com |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Chromatography, High Pressure Liquid
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methods* Escherichia coli / chemistry Plasminogen Activator Inhibitor 1 / analysis* Recombinant Proteins / analysis Sensitivity and Specificity |
| Chemical | |
Reg. No./Substance:
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0/Plasminogen Activator Inhibitor 1; 0/Recombinant Proteins |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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