Size characterization of green fluorescent protein inclusion bodies in E. coli using asymmetrical flow field-flow fractionation?multi-angle light scattering
Size characterization of green fluorescent protein inclusion bodies in E. coli using asymmetrical flow field-flow fractionation?multi-angle light scattering
Available online 18 January 2006.
Jian Luoa, Mats Leemanb, Andras Ballagic, Anders Elfwingc, Zhiguo Sua, Jan-Christer Jansonc and Karl-Gustav Wahlundb
Journal of Chromatography A
Volume 1120, Issues 1-2 , 7 July 2006
ScienceDirect
Copyright ? 2005 Elsevier B.V. All rights reserved.
aNational Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, P.O. Box 353, Beijing 100080, PR China
bDivision of Technical Analytical Chemistry, Center for Chemistry and Chemical Engineering, Faculty of Engineering LTH, Lund University, P.O. Box 124, SE-221 00 Lund, Sweden
cDepartment of Surface Biotechnology, Uppsala Biomedical Center, Uppsala University, P.O. Box 577, SE-751 23 Uppsala, Sweden
Abstract
The goal of this study was to investigate the applicability of asymmetrical flow field-flow fractionation?multi-angle light scattering (AsFlFFF-MALS) for size analysis of green fluorescent protein inclusion bodies (GFPIBs). The size distributions of GFPIBs prepared by various culture conditions were determined. For GFPIBs prepared at 37 ?C the peak maximum hydrodynamic diameter (dH) first increased and then decreased with the increase of the induction times in the presence of 0.1 and 2 mM isopropyl-?-d-thiogalactoside (IPTG). For GFPIBs prepared at 30 ?C the peak maximum dH was constant at about 700 nm irrespectively of the induction times and IPTG concentrations.
Keywords: Asymmetrical flow field-flow fractionation; Multi-angle light scattering; Root mean square radius; Hydrodynamic diameter; Green fluorescence protein inclusion bodies
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