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Development of a novel reporter gene method for determination of ADCC potency of anti-CD20 monoclonal antibody / 药学学报
Acta Pharmaceutica Sinica ; (12): 94-98, 2015.
Artigo em Chinês | WPRIM | ID: wpr-251811
ABSTRACT
The biological activity of ADCC by anti-CD20 monoclonal antibody was determined by BioGlo™ Luciferase Assay System using Jurkat/NFAT-luc+FcγRIIIa cell line as effector cell and WIL2-S cell line as target cell. The developed method was verified for specificity, precision and accuracy. Anti-CD20 monoclonal antibody showed a dose-response mode by the developed method, and the determination result complied with the following four-parameter equation y = (A-D)/[1 + (X/C)(B)] + D. The optimized parameters of the method were determined including the antibodies diluted concentration (18,000 ng·mL(-1)), dilution rate (15), the ratio of effector cell and target cell (61), and induction time (6 h). The values of eight independent tests have passed a statistical test for curve regression analysis, linear or parallelism, which showed the method possessed good specificity. Four different dilute groups of recovery rates sample were determined for 3 times, and the result showed mean relative potencies of (44.39±3.93)%, (72.74±2.78)%, (128.28±7.01)% and (168.19±2.70)% respectively, with a variation coefficient of less than 10%, and the recoveries of (88.78±7.85)%, (96.99±3.70)%, (102.63±5.61)% and (112.12±1.80)% respectively. A novel reporter gene method for determination of biological activity of ADCC by anti-CD20 monoclonal antibody was successfully developed, which showed strong specificity, good reproducibility and high accuracy, and might be used routinely.
Assuntos
Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Farmacologia / Reprodutibilidade dos Testes / Genes Reporter / Antígenos CD20 / Alergia e Imunologia / Anticorpos Monoclonais Murinos / Rituximab / Citotoxicidade Celular Dependente de Anticorpos Limite: Humanos Idioma: Chinês Revista: Acta Pharmaceutica Sinica Ano de publicação: 2015 Tipo de documento: Artigo

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Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Farmacologia / Reprodutibilidade dos Testes / Genes Reporter / Antígenos CD20 / Alergia e Imunologia / Anticorpos Monoclonais Murinos / Rituximab / Citotoxicidade Celular Dependente de Anticorpos Limite: Humanos Idioma: Chinês Revista: Acta Pharmaceutica Sinica Ano de publicação: 2015 Tipo de documento: Artigo