A family of linear hexamer peptide ligands HWRGWV, HYFKFD and HFRRHL, initially identified for their affinity to the Fc portion of human immunoglobulin G (hIgG), also have potential for use in purification of human immunoglobulins A (hIgA) and M (hIgM). HWRGWV demonstrated the strongest binding affinity to hIgM, followed by hIgA and hIgG, respectively. Affinity chromatographic purification of human IgA, IgG and IgM from complete minimum essential media was achieved at different peptide densities. The results suggested that the improved recovery at higher peptide density due to increased binding affinity was compensated by the decrease in purity for all three immunoglobulins. With these knowledge, affinity chromatographic purification of human IgA and IgM from CHO and human B lymphocyte cell lines as well as one step purification of human IgG, IgA and IgM from Cohn fraction II/III were investigated. Different pretreatment methods were employed in order to improve the purity. After caprylic acid pretreatment or the combination of caprylic acid and polyethylene glycol pretreatment, highly purified (over 95% in purity) hIgG was achieved with 60% in overall yield, which is comparable to chromatographic purification method involving two chromatography steps when proteins are isolated from plasma fractions. HIgA (40%) and hIgM (45%) enriched immunoglobulins with over 95% in purity were obtained as the by-products.

Affinity chromatographic purification of antibodies by small peptide ligand   from different sources

243rd National Meeting (2012)

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