Bioassay·Activity Analysis

Evaluation of drug binding activity of anti-CD79b antibody-vc-MMAE*

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  • 1. National Institutes for Food and Drug Control, NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, NMPA Key Laboratory for Quality Research and Evaluation of Biological Products, Beijing 102629, China;
    2. China Pharmaceutical University, Nanjing 211198, China

Received date: 2022-06-14

  Online published: 2024-06-24

Abstract

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.

Cite this article

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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