TY - JOUR
T1 - Analysis of E. coli K5 capsular polysaccharide heparosan
AU - Ly, Mellisa
AU - Wang, Zhenyu
AU - Laremore, Tatiana N.
AU - Zhang, Fuming
AU - Zhong, Weihong
AU - Pu, Dennis
AU - Zagorevski, Dmitri V.
AU - Dordick, Jonathan S.
AU - Linhardt, Robert J.
N1 - Funding Information:
Acknowledgements The authors are grateful to the National Institutes of Health for supporting this research with grants GM38060 and HL096972.
PY - 2011/1
Y1 - 2011/1
N2 - 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 (MN), weight-averaged MWs (M W), and MW ranges of 3,000 to 150,000 Da.
AB - 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 (MN), weight-averaged MWs (M W), and MW ranges of 3,000 to 150,000 Da.
UR - https://www.scopus.com/pages/publications/79251552548
UR - https://www.scopus.com/pages/publications/79251552548#tab=citedBy
U2 - 10.1007/s00216-010-3679-7
DO - 10.1007/s00216-010-3679-7
M3 - Review article
C2 - 20407891
AN - SCOPUS:79251552548
SN - 1618-2642
VL - 399
SP - 737
EP - 745
JO - Analytical and Bioanalytical Chemistry
JF - Analytical and Bioanalytical Chemistry
IS - 2
ER -