In vivo study of the immunogenic properties of the recombinant cellulose-binding domain of Anaerocellum thermophilum

Abstract

Development of novel technologies enabling the one-stage production of preparations of a required degree of purity is one of the most important goals of contemporary medical biotechnology. The incorporation of affinity domains capable of binding to relevant sorbents into recombinant proteins is one of the most promising approaches. The method of one-stage production of protein-cellulose injectants without using chemical conjugates is based on the construction of two-component proteins containing a target antigen and a cellulose-binding domain (CBD) capable of spontaneous binding to cellulose-containing sorbents. To study the immunogenic properties of the CBD, as well as the effect of cellulose sorbents on the immune response, a recombinant CBD was obtained and purified using the one-stage method based on the CBD immobilization on amorphous cellulose; the free CBD protein was isolated. A comparative study of the immunogenic properties of the CBD, the CBD-cellulose complex and the cellulose sorbent was performed using a rat model. The titers of specific antibodies, as well as the spectrum and concentrations of cytokines in the blood serum of rats induced by the obtained preparations, were determined using the BioPlex suspension array system. The effect of the CBD on the induction of cytokine synthesis observed on injection of the free protein and its immobilized form was studied. It has been demonstrated that the immobilization of CBD on the cellulose sorbent enhances the synthesis of specific antibodies. It was found that cellulose immunosorbent is not immunotolerant and induces the synthesis of the cytokines involved in the regulation of the humoral immune response.