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Purification of α2-macroglobulin From Cohn Fraction IV by Immobilized Metal Affinity Chromatography: A Promising Method for the Better Utilization of Plasma

Purification of α2-macroglobulin From Cohn Fraction IV by Immobilized Metal Affinity Chromatography: A Promising Method for the Better Utilization of Plasma

Chaoji Huangfu, Yuyuan Ma, Maomin Lv, Junting Jia, Xiong Zhao, Jingang Zhang

J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Jul 1;1025:68-75.

PMID: 27214605

Abstract:

As an abundant plasma protein, α2-macroglobulin (α2-M) participates widely in physiological and pathological activities including coagulation regulation, antitumor activities, and regulation of cytokines. It also presents a therapeutic potential for radiation injury. A two-step isolation method for the purification of α2-M from Cohn Fraction IV is described. This process includes a salting-out method and immobilized metal affinity chromatography. The LC-ESI-MS/MS analysis and a comparison of the amino acid composition demonstrated that the final product was α2-M. The final protein, with a purity of approximately 95% and a yield of nearly 45%, was obtained from Cohn Fraction IV regardless of plasma haptoglobin type, although all but type 1-1 have previously been considered unfavorable for α2-M preparation. The effects of temperature, pH, and methylamine on α2-M activity were evaluated to avoid activity loss during preparation and preservation. The results suggested that α2-M activity could be readily inactivated at temperatures above 50°C, at pH levels above 9.0 or below 4.0, or in the presence of methylamine. Cohn Fraction IV is usually discarded as a biological waste product in the human serum albumin production process; because the simple process developed in this study is relatively inexpensive, the preparation of α2-M from Cohn Fraction IV may better utilize human plasma, a valuable resource.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR424965	α2-Macroglobulin from human plasma			Price

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