基于USP通则129建立治疗性重组单抗理化特性通用分析平台

目的 基于USP通则129建立针对治疗性单克隆抗体的理化特性通用分析平台。方法 该平台包括分子排阻色谱法、十二烷基硫酸钠-聚丙烯酰胺毛细管电泳法、N-糖的毛细管电泳法、高效液相测定法以及唾液酸离子色谱法。并在此基础上，新增了全柱成像毛细管等电聚焦电泳分析电荷变异体方法，补充了更加灵敏、稳定的超高效液相色谱测定唾液酸含量法。这些方法可用于分析单抗药物的高分子量物质和低分子量物质含量、游离N-糖、唾液酸含量以及电荷变异体测定。结果 通过系统适用性试验和阿达木单抗、贝伐珠单抗、托珠单抗、尼妥珠单抗等4种单抗的应用研究，证明该平台通用性良好。结论 该方法可作为单抗质量控制参考的平台方法，为工业界及监管部门提供参考。

Establishment of Analytic Platform of Physicochemical Characteristics for Therapeutic Monoclonal Antibodies Based on USP General Principle <129>

OBJECTIVE To establish therapeutic monoclonal antibodies(mAbs) drugs' physical and chemical platform based on USP general principle <129>. METHODS This platform was consisted of SEC-HPLC, CE-SDS, CE and HPLC for the analysis of N-glycosylation, and ion chromatography for the evaluation of sialic acid. To improve the platform, a WCID-CIEF method was established as a charge heterogeneity analysis method, and a UPLC method, which was more sensitive and stable, was investigated for the determination of sialic acid. These methods could reveal high molecular weight and low molecular weight variants, N-glycosylation, sialic acid content and charge variants of mAb drugs. RESULTS These methods were verified through system suitability tests and applied in four mAbs, including Adalimumab, Bevacizumab, Tocilizumab, Nimotuzumab. The results showed that these platform methods were suitable for the physical and chemical properties analysis of mAb drugs. CONCLUSION This platform may provide reference methods in mAb drugs' quality control for industry and regulators.