山西医科大学学报2025，Vol.56Issue(12)：1416-1423,8.DOI:10.13753/j.issn.1007-6611.2025.12.013
抗IL-17A单克隆抗体药物的质量控制
Quality control of anti-IL-17A monoclonal antibody drug
摘要
Abstract
Objective To establish quality control methods for key quality attributes(KQAs)of anti-interleukin 17A(IL-17A)mono-clonal antibodies(mAbs).Methods The biological activity was determined by GROα cytokine bioactivity assay.Peptide mapping was analyzed by reverse-phase chromatography.The isoelectric point was measured by capillary isoelectric focusing electrophoresis(cIEF).Purity was evaluated by reduced/non-reduced capillary electrophoresis-sodium dodecyl sulfonate(CE-SDS)and size exclusion-high performance liquid chromatography(SEC-HPLC).Charge heterogeneity was assessed by ion exchange chromatography(IEC-HPLC).Glycosylation of anti-IL-17 mAb was analyzed by hydrophilic-ultra high performance liquid chromatography(HILIC-UPLC).The binding activity of IL-17A was detected by ELISA.Results The half maximal effective concentration(EC50)for the biological activity of anti-IL-17A monoclonal antibodies was(6.70±1.19)ng/mL.The EC50 of IL-17A binding activity was(35.60±3.70)ng/mL.The main peak isoelectric point determined by cIEF was 7.30±0.004.The percentage of main peak area by non-reduced CE-SDS was(96.95±0.12)%,and the sum of peak area percentages of heavy chain(HC)and light chain(LC)in reduced CE-SDS was(98.38±0.11)%.The percentage of main peak area by SEC-HPLC was(99.55±0.01)%.In ion exchange chromatography,the percentage of main peak area was(73.07±0.12)%,the percentage of acidic peak area was(16.52±0.05)%,and the percentage of basic peak area was(10.41±0.10)%.In N-glycan analysis,the total proportion of high-mannose glycosylation 5 monosaccharides was(1.96±0.03)%,the total proportion of fucosylation was(88.69±0.44)%,and the total proportion of galactosylation monosaccharides was(11.96±0.03)%.Conclusion Quality control analytical methods for the critical quality attributes(CQAs)of the anti-IL-17A monoclonal anti-body have been successfully established,and can be applied to the quality control of this category of monoclonal antibody products.关键词
抗白介素17A单抗/白介素17A/纯度/生物学活性/电荷异质性/质量控制Key words
anti-interleukin 17A monoclonal antibody/interleukin 17A/purity/biological activity/charge heterogeneity/quality control分类
医药卫生引用本文复制引用
CUI Chunbo,DUAN Maoqin,WANG Lan,YU Chuanfei,NI Yongbo,WU Gang,LI Meng,LIU Chunyu,DU Jialiang,CUI Yongfei,GUO Luyun,YANG Yalan..抗IL-17A单克隆抗体药物的质量控制[J].山西医科大学学报,2025,56(12):1416-1423,8.基金项目
国家重点研发计划项目(2021YFF0600804) （2021YFF0600804）
