A new purification process for goose immunoglobulin IgY(ΔFc) with hydrophobic charge-induction chromatography

Abstract

Hydrophobic charge-induction chromatography (HCIC) is a novel bioseparation technique, especially for antibody purification. In this study, HCIC was used for the purification of goose immunoglobulin IgY(ΔFc). IgY(ΔFc) is a kind of unique immunoglobulin existing in waterfowl, which natively lacks C H3 and C H4 domains of heavy chains and has some particular therapeutic applications. A new HCIC ligand, 2-mercapto-1-methyl-imidazole (MMI), was coupled to the divinyl-sulfone-activated agarose matrix to prepare the HCIC gel. The adsorption isotherms of IgY(ΔFc) were investigated at different pHs. The saturated capacity of IgY(ΔFc) at pH 5.0 could reach 187.5 mg/ml gel, and the pH-dependent adsorption behaviors were found. After the goose plasma was pre-treated with caprylic acid to precipitate some impurities, the supernatant could be directly loaded onto the HCIC column for IgY(ΔFc) separation. The operation conditions were optimized, including the loading pH, elution pH and the loading volume. High separation efficiency was obtained with final purity of 98.6% and the yield of 85.0%. The results indicate that the new process with HCIC can be a promising technology for the cost-effective separation of IgY(ΔFc) from plasma.