APPLICATION OF COVALENT AFFINITY-CHROMATOGRAPHY FOR SEPARATION OF ZN-THIONEIN FROM LIVER OF RATS EXPOSED TO ZN

Covalent affinity chromatography with thiol-disulphide interchange (CA C-TDI) was employed for separation of Zn-thionein from the liver of Zn induced white rats (20.0 mg of Zn per kg body weight). Initial purifi cation of Zn-thionein was obtained by gel permeation chromatography on Sephadex G-200 gel. Two-, three- and four-step elutions were used for separation of Zn-thionein by CAC-TDI method resulted in dissociation of Zn from Zn-thionein with appearing of monomers, oligoaggregates and polyaggregates of separated metallothionein as inferred from rechroma tography by gel permeation chromatography on Sephadex G-200. The Zn ap peared as a low molecular weight complexes with TNB about 2.0-3.0 kDa.