Doneanu Catalin E, Chen Weibin
Waters Corporation, 34 Maple Street, Milford, MA, 01757, USA,
Methods Mol Biol. 2014;1129:341-50. doi: 10.1007/978-1-62703-977-2_25.
A generic method for the identification and quantification of host-cell proteins (HCPs) in protein biopharmaceuticals is described. Therapeutic proteins and HCPs were converted to complex peptide mixtures following tryptic digestion. Comprehensive peptide separations were performed using online two-dimensional capillary liquid chromatography-(LC) involving high-pH reversed phase (RP)/low-pH RP separations. We applied this method to the analysis of HCP impurities in monoclonal antibody (mAb) preparations.
描述了一种用于鉴定和定量蛋白质生物制药中宿主细胞蛋白（HCP）的通用方法。治疗性蛋白质和HCP在胰蛋白酶消化后转化为复杂的肽混合物。使用在线二维毛细管液相色谱法（LC）进行全面的肽分离，该方法涉及高pH反相（RP）/低pH RP分离。我们将此方法应用于单克隆抗体（mAb）制剂中HCP杂质的分析。
