Membrane affinity chromatography for analysis and purification of biopolymers |
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Authors: | Dongmei Zhou Hanfa Zou Jianyi Ni Hailin Wang Li Yang Yukui Zhang |
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Institution: | (1) National Chromatographic R & A Center, Dalian Institute of Chemical Physics, The Chinese Academy of Sciences, 116011 Dalian, P. R. China |
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Abstract: | Summary Affinity columns suitable for HPLC were prepared by immobilization of various ligands of protein A, human IgG, human IgM and
pectinase on GMA modified cellulose membrane. The adsorption capacity, affinity efficiency and activity recovery of various
IgGs on these affinity columns were measured. It was observed that the length of the coupling arm plays a very important role
in affinity efficiency, and the effect of eluent flow-rate on adsorption capacity was very small. The protein A column was
exploited for the process monitoring of dog IgG in clinical experiments on immuno-adsorption therapy. A pectinase column was
used for the determination of polygalacturonase inhibiting proteins first purified on a hydroxyapatite column. It took only
about 2.5 min for analysis at a flow-rate of 1.0 mL min−1. The high speed analysis of biopolymers could be performed at a flow rate of 6.0 mL min−1 within 15 s. Membrane affinity chromatography gives good reproducibility, high efficiency, low column-pressure and is rapid.
It can also be used for micro-scale purification of biopolymers. |
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Keywords: | Column liquid chromatography Membrane stationary phase Immunoaffinity analysis Affinity efficiency Adsorption capacity |
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