Objective: To study the binding activity of anti-CD79b antibody and anti-CD79b antibody-vc- MMAE by establishing surface plasmon resonance(SPR) and enzyme linked immunosorbent assay(ELISA) measures. Methods: In this experiment, SPR and Protein A chip were used to determine the kinetic constants, including binding constant(Ka), dissociation constant (Kd), affinity constant(KD), half maximal effective concentration(EC50) and relative potency between anti-CD79b antibody and anti-CD79b antibody-vc-MMAE and their target CD79b protein. Classical ELISA was used to determine the EC50 and relative potency of anti-CD79b antibody and anti-CD79b antibody-vc-MMAE with CD79b. Results: After three experiments, the kinetic and affinity parameters between anti-CD79b antibody and CD79b include: Ka, Kd and KD, which were (2.61±0.14)×106 L·mol-1·s-1, (1.87±0.12)×10-2 s-1, (7.17±0.30)×10-9mol·L-1 respectively, RSDs were<10%. The kinetic and affinity parameters between anti-CD79b antibody-vc-MMAE and CD79b include: Ka, Kd and KD, which were(2.99±0.26)×106 L·mol-1·s-1, (1.46±0.04)×10-2 s-1, (4.88±0.31)×10-9 mol·L-1 respectively, RSDs were<10%. After three experiments, the EC50 and relative potency between anti-CD79b antibody and CD79b determined by SPR were (78.68±6.35) ng·mL-1, (84.62±6.63)% respectively, RSDs were<10%. The EC50 and relative potency between anti-CD79b antibody-vc-MMAE and CD79b determined by SPR were (35.93±0.75) ng·mL-1, (84.74±1.76)% respectively, RSDs were<5%. The EC50 and relative potency between anti-CD79b antibody and CD79b determined by ELISA were (22.66±0.41) ng·mL-1, (95.70±1.74)% respectively, RSDs were<5%. The EC50 and relative potency between anti-CD79b antibody-vc-MMAE and CD79b determined by ELISA were (69.19±1.71) ng·mL-1, (97.64±2.39)% respectively, RSDs were <5%. The results of SPR and ELISA were basically consistent. Conclusion: In this study, the kinetic and affinity parameters of anti-CD79b antibody and anti-CD79b antibody-vc-MMAE against target binding were determined by SPR.The EC50 and relative potency analyzed by SPR and ELISA are compared. The study provides a reference for the development of this kind of ADC drugs.
LI Meng, ZHAO Xue-yu, YU Xiao-juan, YANG Ya-lan, LONG Cai-feng, YU Chuan-fei, WANG Lan
. Evaluation of drug binding activity of anti-CD79b antibody-vc-MMAE*[J]. Chinese Journal of Pharmaceutical Analysis, 2022
, 42(10)
: 1754
-1762
.
DOI: 10.16155/j.0254-1793.2022.10.08
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