Isolation of Fibrinogen Aα-Chain by Affinity Chromatography on Concanavalin A-Sepharose

Summary

A method was developed for isolation of the Aa-chain from S-carboxamidomethylated fibrinogen. A mixture of the three constituent polypeptide chains of human fibrinogen was applied onto a column of concanavalin A-Sepharose. While the carbohydrate-free Aα-chain was not delayed on the affinity chromatography column, both glycosylated subunit chains, Bβ- and γ-chain, were adsorbed to the insolubilized lectin and were quantitatively eluted from the column with 0.2 M methyl-α-D-mannoside. SDS- electrophoresis on polyacrylamide gel was employed for analysis of chromatographic fractions. Complete recovery of the Aα-chain was observed. The described procedure is very simple and permits isolation of large amounts of pure Aα-chain from S-carboxamidomethylated fibrinogen.

Keywords

Fibrinogen - Aα chain - Concanavalin A - Affinity chromatography

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