Article
MA Chun-mei1,2,3, HUANG Xiao-xing, CAO Zhong-lian, ZHENG Bin, MENG Li-zheng, LIU Li*
Objective To verify the biological activity of adalimumab by using Biacore T200 system. Methods Adalimumab was immobilized to the CM5 biosensor chip, and the dynamics constants of recombinant human tumor necrosis factor-α (TNF-α) and recombinant cynomolgus TNF-α were obtained. To verify species difference, the binding values of different species of recombinant TNF-α were measured. The median effective concentration of adalimumab for inhibiting the toxicity of recombinant TNF-α to L929 cell was measured. Results The dynamics constants of adalimumab binded to recombinant human TNF-α Ka was (2.114±0.007)×105; Kd was (5.315±0.220)×10-5 and KD was (2.515±0.107)×10-10. The dynamics constants of adalimumab binded to recombinant cynomolgus TNF-α Ka was (2.574±0.068)×105; Kd was (1.554±0.124)×10-5 and KD was (5.320±1.271)×10-10. The binding values of recombinant human TNF-α, recombinant cynomolgus TNF-α, recombinant mouse TNF-α and recombinant rat TNF-α were 84.3, 66.4, 1.3 and -0.8 RU, respectively. The median effective concentration of the adalimumab inhibitory effect on recombinant human TNF-α and recombinant cynomolgus TNF-α cell toxicity were 194.0 pmol/L and 685.5 pmol/L respectively. However, it had no influence on recombinant mouse TNF-α and recombinant rat TNF-α cell toxicity. Conclusion Biacore T200 system is simple, rapid, accurate and suitable for the biological activity test and the pharmacodynamic evaluation of recombinant human TNF-α monoclonal antibody.