Therapeutic efficacy of three bispecific antibodies on rheumatoid arthritis mice models / 药学学报

In order to obtain the lead compound for treatment of rheumatoid arthritis (RA), in this study, therapeutic efficacy of three bispecific antibodies (BsAB-1, BsAB-2 and BsAB-3) against both hIL-1beta and hIL-17 were compared on CIA model mice. First, by ELISA method we compared the binding capacity of the three bispecific antibodies to the two antigens. The results showed that all three antibodies could simultaneously bind both antigens, among these antibodies, BsAB-1 was superior over BsAB-2 and BsAB-3. CIA model was established with chicken type II collagen (CII) and developed RA-like symptoms such as ankle swelling, skin tight, hind foot skin hyperemia. The CIA mice were treated with three antibodies once every two days for total of 29 days. Compared with the CIA model mice, the RA-like symptoms of the antibody treated-mice significantly relieved, while the BsAB-1 treated-mice were almost recovered. CII antibody level in the serum and cytokines (IL-2, IL-1beta, IL-17A and TNF-alpha) expression in the spleen were examined. Compared with the CIA model mice, all three antibodies could significantly reduce CII antibody and cytokine expression levels. BsAB-1 antibody was more potent than BsAB-2 and BsAB-3. In summary, BsAB-1 is superior over BsAB-2 and BsAB-3 in amelioration of RA symptoms and regulation of CII antibody production and pro-inflammatory cytokine expression, therefore, BsAB-1 can be chosen as a lead compound for further development of drug candidate for treatment of RA.

Construction, expression, and bio-activity assay of an anti-IL-1βscfv and TNFR1 fusion protein / 中华微生物学和免疫学杂志

Objective To express the anti-IL-1βscfv and soluble TNF receptor 1 (sTNFR1),and analyze their bio-activities.Methods sTNFR1 was obtained by RT-PCR from the total RNA of HeLa cells,and fused with IL-1βscfv by the hinge fragment of IgG molecule.The fusion gene IL-1scfv:TNFR1 was cloned into the expression vector pET27b(+).The fusion protein was expressed and purified from inclusion bodies.Results The ELISA analysis showed that the fusion protein could bind hIL-1β and hTNF-α respectively in a dose-dependent manner,indicating that scfv and sTNFR in the fusion protein can form the correct spatial configuration.The dolt-blot analysis showed that the fusion protein could concurrently bind with hIL-1β and hTNF-α,indicating that the combination of the two parts of the fusion protein does not influence each other for binding to their target molecules.The bioactivity assay showed that the fusion protein could inhibit both the cytotoxicity of hTNF-α on L929 cells and hIL-1β-induced proliferation of L929 cells,indicating that the fusion protein has the ability to neutralize hTNF-α and hIL-1β.Conclusion A bispecific fusion protein IL-1scfv:TNFR1 was successfully constructed.The fusion protein has the ability to inhibit the biological activity of hTNF-α and hIL-1β,and provides a drug candidate for the treatment of rheumatoid arthritis.