Preparation, characterization and immunostimulatory effects of CRD2 and CRD3 from TNF receptor-1 encapsulated into pegylated liposomal nanoparticles

Tumor necrosis factor receptor 1 (TNFR1) is expressed on tumor cells and induces cell apoptosis through triggering proapoptotic cascade in these cells. Agonist antibodies specifc for TNFR1 mimic the role of receptor ligands and can induce apoptosis in tumor cells. This study aims to prepare cysteine-rich domain2 (CRD2) and CRD3 of the extracellular region of TNFR1 receptor as a recombinant protein, and encapsulate the protein into pegylated liposomal nanoparticles to obtain an adjuvant antigen for cancer treatment. In this study, the size, size distribution and zeta potential of particles were determined by Zetasizer, and their stability was evaluated after 3 months of their production time. Moreover, the adjuvant efect of liposomal nanoparticles was evaluated in an in vitro environment using dendritic and lymphocyte cells. The size and zeta potential of protein-loaded pegylated liposomal nanoparticles were 274±13 nm and − 28 mV, respectively, with 78%±2.3 protein encapsulation efciency. In addition, the adjuvant efect of liposomal nanoparticles as the recombinant protein carrier was confrmed, in which the lymphocyte proliferation was increased by 1.5-fold in the presence of liposomal nanoparticles. The results of the present study suggest that CRD2 and CRD3 encapsulated into pegylated liposomal nanoparticles can be used as adjuvant antigens for the production of specifc agonist antibody for TNFR1 stimulation in an in vivo environment.