Journal
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 399, Issue 2, Pages 737-745Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-010-3679-7
Keywords
E. coli K5 capsular polysaccharide; Heparosan; Molecular weight determination; PAGE; ESI-FT-MS; Continuous-elution preparative electrophoresis
Funding
- National Institutes of Health [GM38060, HL096972]
- NATIONAL HEART, LUNG, AND BLOOD INSTITUTE [R01HL096972] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [R01GM038060] Funding Source: NIH RePORTER
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Heparosan is the key precursor for the preparation of bioengineered heparin, a potential replacement for porcine intestinal heparin, an important anticoagulant drug. The molecular weight (MW) distribution of heparosan produced by the fermentation of E. coli K5 was investigated. Large-slab isocratic and mini-slab gradient polyacrylamide gel electrophoresis (PAGE) were used to analyze the MW and polydispersity of heparosan. A preparative method that allowed fractionation by continuous-elution PAGE was used to obtain heparosan MW standards. The MWs of the heparosan standards were determined by electrospray ionization Fourier-transform mass spectrometry (ESI-FT-MS). A ladder of the standards was then used to determine the MW properties of polydisperse heparosan samples. Unbleached and bleached heparosan produced by fermentation of E. coli K5 had similar number-averaged MWs (M-N), weight-averaged MWs (M-W), and MW ranges of 3,000 to 150,000 Da.
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