Affinity Chromatography Methods and Protocols 3rd Ed.

Antibody Purification from Human Plasma by Metal-Chelated Affinity Membranes

Handan Yavuz, Nilay Bereli, Fatma Yılmaz, Canan Armutcu, and Adil Denizli

Abstract

Immobilized metal ion affinity chromatography (IMAC) has been used for purification of proteins. IMAC introduces a new approach for selectively interacting biomolecules on the basis of their affinities for metal ions. The separation is based on different binding abilities of the proteins to the chelated metal ions on support. Here, N-methacryloyl-(L)-histidine methyl ester (MAH) is used as the metal-chelating ligand. Poly(hydroxyethyl methacrylate) Poly(HEMA) based membranes were prepared by photo-polymerization technique. Then, Zn2+, Ni2+, Co2+, and Cu2+ ions were chelated directly on the poly(HEMA-MAH) membranes for purification of immunoglobulin G (IgG) from human plasma.

Molecularly Imprinted Cryogels for Human Serum Albumin Depletion

Muge Andac, Igor Yu Galaev, Handan Yavuz, and Adil Denizli

Abstract

Molecularly imprinted polymers can be used for the selective capture of a target molecule from complex medium. Cryogels novel matrices, which characterized by their supermacropores that makes their use advantageous when studying with biological samples. By combining high selectivity of the molecular imprinting approach with using cryogel as a base polymer, in this protocol, preparation of the albumin- imprinted cryogels is described. This material is a useful candidate for the selective albumin depletion from the human serum sample prior to the detailed proteomic analysis.