全人源化抗人TNF-α单克隆抗体毛细管区带电泳鉴别方法的建立及验证

发布时间：2018-07-07 22:26

本文选题：人肿瘤坏死因子α + 单克隆抗体　；参考：《微生物学免疫学进展》2016年06期

【摘要】：目的建立及验证用于全人源化抗人TNF-α(Tumor necrosis factor-α)单克隆抗体(简称抗人TNF-α单抗)鉴别的毛细管区带电泳(Capillary zone electrophoresis,CZE)方法。方法采用DB-1毛细管,以样品检测分离度、迁移时间、电流及峰形为判断标准,筛选CZE过程中的各种参数(缓冲液p H、ε-氨基乙酸浓度、乙酸钠浓度、羟丙基纤维素浓度、样品稀释剂、进样时间及运行电压等),建立CZE方法,并对方法的专属性、精密度、中间精密度进行验证。结果建立的CZE方法参数条件为:缓冲液p H为5.0,EACA浓度为300 mmol/L,乙酸钠浓度为20 mmol/L,HPMC质量分数为0.05%,用50 mmol/L Tris稀释样品,进样时间为15 s,运行电压为20 k V。样品制备重复性主峰迁移时间RSD为0.459%,中间精密度RSD为1.145%。结论 CZE方法分离度高、专属性强、重复性和精密度好、简单快速,可作为全人源化抗人TNF-α单抗的鉴别方法。
[Abstract]:Objective to establish and verify a capillary zone electrophoresistics (CZE) method for the identification of total human necrosis factor- 伪 monoclonal antibodies (McAbs) against human TNF- 伪. Methods DB-1 capillary tube was used to screen the parameters (buffer pH, 蔚 -aminoacetic acid concentration, sodium acetate concentration, hydroxypropyl cellulose concentration) in the CZE process according to the criteria of sample separation, migration time, current and peak shape. Sample diluent, sample injection time and operation voltage), establish CZE method, and verify the specificity, precision and intermediate precision of the method. Results the parameters of CZE method were as follows: buffer pH = 5.0 渭 mol / L, sodium acetate concentration = 20 mmol / L, mass fraction of HPMC = 0.05. The sample was diluted with 50 mmol / L Tris, the injection time was 15 s, and the operating voltage was 20 kV. The reproducibility of the main peak migration time was 0.459, and the intermediate precision RSD was 1.145. Conclusion the CZE method has the advantages of high separation degree, strong specificity, good repeatability and precision, simple and rapid, and can be used as the identification method of total humanized anti human TNF- 伪 monoclonal antibody.
【作者单位】：湖北医药学院附属襄阳医院超声科;武汉生物制品研究所有限责任公司抗体研究室;
【基金】：国家科技重大专项课题资助项目(2011ZX09506-005)
【分类号】：R392

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