Protein Expression and Purification 1997-02-01

Production of biologically active recombinant avidin in baculovirus-infected insect cells.

K J Airenne, C Oker-Blom, V S Marjomäki, E A Bayer, M Wilchek, M S Kulomaa

Index: Protein Expr. Purif. 9(1) , 100-8, (1997)

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Abstract

An efficient lepidopteran insect cell system was established for the expression of a recombinant form of chicken egg-white avidin. The gene product was obtained in both secreted and intracellular forms, and biologically active recombinant avidin was isolated using affinity chromatography on an iminobiotin-agarose column. Similar to the known quaternary structure of the native egg-white protein, the purified recombinant protein was glycosylated and assembled mainly into tetramers. Like native avidin, the recombinant tetramer also exhibited a high level of thermostability, and was further stabilized upon binding biotin. The biotin-binding and structural properties of the recombinant avidin are thus similar to those of the natural egg-white protein, and the insect system is appropriate both for future site-directed mutagenesis studies and for the production of avidin fusion proteins.