Purification and separation of mannan-binding lectin (MBL) and MBL-associated serine proteases complex from human plasma

Abstract

Abstract: Objective To separate mannan-binding lectin (MBL) and MBL-associated serine proteases (MASPs) from human plasma. Methods A two-step affinity chromatography on underivatized sepharose 4B was employed for purification of MBL-MASP complex, followed by gel filtration on a Sephacryl S-300 column for separation of MBL and MASPs from the complex. The purification procedures were performed at 4℃ with the addition of two proteolytic inhibitors, phenyl methylsulfonyl fluoride and 1,10-phenanthroline during affinity chromatography but not in the gel filtration buffer. Results Preparations of highly purified MBL and proenzyme MASPs were obtained. The purified MBL was shown by SDS-PAGE and Western blotting to be a functional multimer composed of 28 000 and 32 000 peptide chains, with high bioactivity as demonstrated by ligand-binding assay and yeast agglutination experiment. Conclusion A simple and convenient procedure is established successfully for the purification of MBL and the proenzyme MASPs.

CLC Number:

R392-33

CHEN Yue, ZHANG Li-yun, CHEN Zheng-liang. Purification and separation of mannan-binding lectin (MBL) and MBL-associated serine proteases complex from human plasma[J]. Journal of Southern Medical University, 2004, 24(12): 1373-1377.

References

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