Capillary zone electrophoresis tandem mass spectrometry detects low concentration host cell impurities in monoclonal antibodies. Issue 4 (February 2016)

• Record Type: Journal Article
• Authors: Zhu, Guijie; Sun, Liangliang; Heidbrink‐Thompson, Jennifer; Kuntumalla, Srilatha; Lin, Hung‐yu; Larkin, Christopher J.; McGivney, James B.; Dovichi, Norman J.
• Is Part Of: Electrophoresis. Volume 37:Issue 4(2016)
• Page Start: 616
• Page End: 622
• Publication Date: 2016-02
• Subjects: Bottom‐up proteomics -- CZE -- Host cell proteins; Electrophoresis -- Periodicals; Electrophoresis -- Periodicals; 541.372
• DOI: 10.1002/elps.201500301
• Languages: English
• ISSNs: 0173-0835

Abstract

Abstract : We have evaluated CZE‐ESI‐MS/MS for detection of trace amounts of host cell protein impurities in recombinant therapeutics. Compared to previously published procedures, we have optimized the buffer pH used in the formation of a pH junction to increase injection volume. We also prepared a 5‐point calibration curve by spiking 12 standard proteins into a solution of a human mAb. A custom CZE‐MS/MS system was used to analyze the tryptic digest of this mixture without depletion of the antibody. CZE generated a ∼70‐min separation window (∼90‐min total analysis duration) and ∼300‐peak capacity. We also analyzed the sample using ultra‐performance LC‐MS/MS. CZE‐MS/MS generated approximately five times higher base peak intensity and more peptide identifications for low‐level spiked proteins. Both methods detected all proteins spiked at ∼100 ppm level with respect to the antibody.