基因工程药物中宿主细胞蛋白的检测与控制  

Detection and control of host cell proteins in genetic engineering pharmaceuticals

作　　者：王志明[1] 

机构地区：[1]华北制药集团新药研究开发有限责任公司抗体药物研制国家重点实验室,石家庄050015

出　　处：《中国新药杂志》2016年第22期2550-2557,共8页Chinese Journal of New Drugs

基　　金：国家"重大新药创制"科技重大专项资助项目(2012ZX09101313)

摘　　要：宿主细胞蛋白(host cell proteins,HCPs)是基因工程菌株或细胞株自身产生的与基因工程目的产物无关的蛋白混合物,其中有些蛋白是工程菌株或细胞株生存、繁殖及其他正常生理活动所必须的,其在产品生产过程与目的产物共同纯化且不能被去除。HCP的组成与丰度在生产的各个阶段与最终产品都各自不同,其决定于许多不同因素。ELISA(enzyme-linked-immunosorbent assay)是HCP定量检测中最常用的分析手段,但是由于依赖于特异性抗体的产生,在许多情况下也限制了该方法的检测应用。可视化检测技术(如双向差异凝胶电泳)与定性检测技术(如质谱分析)成为ELISA检测的有力验证与补充。对HCP及其产生机制越来越深入的理解有助于我们设计基因工程药物的生产过程(从上游细胞系的选择到下游纯化手段的选择),从而降低或去除最终产品中的HCP。Host cell proteins （HCPs） are those produced or encoded by the organisms and unrelated to the intended recombinant product. Some are necessary for growth, survival and normal cellular processing, whereas others may be non-essential, simply carried along as baggage. Though commonly present in .small quantities （parts per million expressed as nanograms per milligrams of the intended recombinant protein）, HCP is a critical quality attribute that affects patient safety, so much effort and cost are expended by industry to remove them. The composition and abundance of HCPs present in various steps of manufacturing processes and in the final drug substance depend on many factors. An enzyme-linked-immunosorbent assay （ELISA） is the workhorse method for HCP testing due to its high throughput, sensitivity and selectivity, however dependence on the production of anti-HCP antibodies, thus there are many limitations of the applications. Alternative approaches are also now emerging, such as 2D-DIGE and MS, which complement ELISA and allow the detection, identification and monitoring of specific HCPs. We anticipate that more fundamental understanding of impurities and the product itself would now be applied to upstream processing design （cell line selection and so on） , and to downstream processing development. These will reduce or remove general and specific problematic HCPs before and during downstream processing.

关 键 词：宿主细胞蛋白 组成和丰度 酶联免疫吸附检测 双向差异凝胶电泳 质谱 细胞工程 

分 类 号：R978.7[医药卫生—药品]