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Design of suitable carrier buffer for free‐flow zone electrophoresis by charge‐to‐mass ratio and band broadening analysis
Author(s) -
Kong Fanzhi,
Yang Ying,
He Yuchen,
Zhang Qiang,
Li Guoqing,
Fan Liuyin,
Xiao Hua,
Li Shan,
Cao Chengxi
Publication year - 2016
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201600040
Subject(s) - electrophoresis , analytical chemistry (journal) , chemistry , phycobiliprotein , capillary electrophoresis , chromatography , materials science , cyanobacteria , biology , bacteria , genetics
In this work, charge‐to‐mass ratio (C/M) and band broadening analyses were combined to provide better guidance for the design of free‐flow zone electrophoresis carrier buffer (CB). First, the C/M analyses of hemoglobin and C‐phycocyanin (C‐PC) under different pH were performed by CLC Protein Workbench software. Second, band dispersion due to the initial bandwidth, diffusion, and hydrodynamic broadening were discussed, respectively. Based on the analyses of the C/M and band broadening, a better guidance for preparation of free‐flow zone electrophoresis CB was obtained. Series of experiments were performed to validate the proposed method. The experimental data showed high accordance with our prediction allowing the CB to be prepared easily with our proposed method. To further evaluate this method, C‐PC was purified from crude extracts of Spirulina platensis with the selected separation condition. Results showed that C‐PC was well separated from other phycobiliproteins that have similar physicochemical properties, and analytical grade product with purity up to 4.5 (A620/A280) was obtained.