Biopolymers and cell. 2006; 22 (2): 157 - 161

Immobilized single chain antibodies for affinity purification of recombinant human IFN-α2b

P. V. Gilchuk, O. V. Okunev, D. M. Irodov, M. V. Pavlova, O. Ya. Yakovenko

A laboratory method for obtaining immunoaffine media for chromatographic purification of proteins based on orienting and non-covalent immobilization of recombinant antibody fragments on cellulose matrix is described. The single-chain antibody (ScFv) against human IFN-α2b was genetically fused to cellulose-binding domain (CBD) from cellulosome complex of Clostridium thermocellum and expressed in Escherichia coli. After the isolation of the target protein in functionally active form from bacteria cells the directed immobilization on microgranular cellulose has been carried out. The cellulose with immobilized ScFv-CBD-fusion was used as affinity medium to perform the purifications of recombinant human IFN-α2b

Key words: single chain antibodies, immunoaffinity chromatography, protein immobilization, fusion protein.