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Recombinant TNF-Binding Protein from Variola Virus as a Novel Potential TNF Antagonist

I. P. Gileva*, T. S. Nepomnyashchikh, I. A. Ryazankin, and S. N. Shchelkunov

FGUN State Research Center of Virology and Biotechnology Vector, 630559 Koltsovo, Novosibirsk Region, Russia; fax: (383) 336-7409; E-mail: gileva@vector.nsc.ru

* To whom correspondence should be addressed.

Received April 3, 2008; Revision received May 25, 2009
Gel-filtration chromatographic separation of the lysate of Sf21 insect cells infected with recombinant baculovirus BVi67 containing the gene for TNF-binding protein (CrmB) of variola virus (VARV) revealed that hTNF-cytotoxicity neutralization activity is associated with a fraction corresponding mainly to high molecular weight proteins (above 500 kDa) and less with fractions corresponding to proteins of 270 or 90 kDa. The recombinant VARV-CrmB protein has been purified by affinity chromatography. Difference in the experimentally determined and estimated (according to amino acid composition) VARV-CrmB molecular weight is due to glycosylation of the recombinant protein expressed in the insect cells. VARV-CrmB neutralizes in vitro the cytotoxic effect of hTNF and hLTα, and its TNF-neutralizing activity is two to three orders of magnitude higher compared to the analogous effects of type I and II soluble TNF receptors, comparable with the activity of mAb MAK195, and somewhat lower than the effect of the commercial drug Remicade.
KEY WORDS: tumor necrosis factor, TNF-binding protein, variola virus, viroceptors, septic shock

DOI: 10.1134/S0006297909120098