The assessment of TNF-α interaction with oligopeptide Trp-Asn-Trp-Val in vitro

Abstract

The hyperproduction of TNF-α in the human body is a pathogenetic factor of many diseases development. The widespread clinical use of anticytokine drugs based on antibodies has a number of limitations. Therefore, the development of low molecular weight compounds for this cytokine blocking is a priority. Objectives. To experimentally study the interaction of TNF-α with the synthetic oligopeptide Trp-Asn-Trp-Val, which is a structural analog of the soluble TNF-α receptor. Material and methods. For registration of the interaction of the cytokine with oligopepetide the change in the concentration of TNF-α after its binding to the free and immobilized form of the oligopeptide was assessed. The immobilization of the oligopeptide was carried out by adsorption on the bottom of the well of a 96-well plate and incorporation of the peptide into a three-dimensional network of polyacrylamide gel. The concentration of TNF-α was determined by enzyme immunoassay. The results of the study have shown that the dependence of the bound cytokine amount on its initial concentration is hyperbolic. The free form of the oligopeptide binds 6.8 (5.5; 8.1) pM/ml, adsorbed on the plate – 5.1 (4.3; 5.8) pM/ml of TNF-α. Immobilization of the oligopeptide in the gel significantly increases the ability of Trp-Asn-Trp-Val oligopeptide to bind TNF-α. The maximum concentration of TNF-α in blood plasma bound by the immobilized in gel peptide made up 16.8 (14.3; 19.2) pM/ml. Thus, the conducted studies have shown that the oligopeptide Trp-Asn-Trp-Val, both in free and immobilized form, possesses a pronounced ability to bind TNF-α and may be used as a ligand for hemosorbents.

Keywords

• tumor necrosis factor-alpha
• oligopeptide
• cytokines