Comprehensive analysis of host cell impurities in monoclonal antibodies with improved sensitivity by capillary zone electrophoresis mass spectrometry. Issue 3 (14th December 2016)

• Record Type: Journal Article
• Authors: Zhang, Zhenbin; Albanetti, Thomas; Linkous, Travis; Larkin, Christopher J.; Schoner, Ronald; McGivney, James B.; Dovichi, Norman J
• Is Part Of: Electrophoresis. Volume 38:Issue 3/4(2017)
• Page Start: 401
• Page End: 407
• Publication Date: 2016-12-14
• Subjects: Cationic solid phase extraction -- CZE‐MS/MS -- Electrokinetically pumped nanoelectrospray -- Host cell proteins -- pH bumps; Electrophoresis -- Periodicals; Electrophoresis -- Periodicals; 541.372
• DOI: 10.1002/elps.201600390
• Languages: English
• ISSNs: 0173-0835

Abstract

Abstract : Four methods were compared for analysis of host‐cell protein (HCP) impurities in a recombinant mAb. First, CZE‐MS/MS was used to analyze the digest of an HCP sample following extraction of the mAb with proteins A and L affinity columns; 220 protein groups and 976 peptides were identified from the depleted HCP digest. Second, a nanoACQUITY UltraPerformance LCH system was also used to analyze the depleted HCP digest; 34 protein groups and 53 peptides from 50 ng of the depleted HCP digest and 290 protein groups and 1011 peptides were identified from 1 μg of the depleted HCP digest. Third, 185 protein groups and 709 peptides were identified by CZE‐MS/MS from the HCP digest without depletion. Fourth, a strong cation exchange SPE was coupled to CZE‐ESI‐MS/MS using online pH gradient elution for analysis of the HCP digest without depletion. A series of five pH bumps were applied to elute peptides from the strong cation exchange monolith followed by analysis using CZE coupled to a Q Exactive HF mass spectrometer; 230 protein groups and 796 peptides were identified from the HCP digest without depletion.