Capillary electrophoresis-mass spectrometry as a characterization tool for therapeutic proteins.
With the increasing use of capillary electrophoresis (CE) in the biotechnology industry, there is a demand for analytical tools and methodology that can be used to characterize CE profiles. This article describes the implementation and optimization of a robust online CE-mass spectrometry (CE-MS) system used for the characterization of several CE assays developed at Genentech Inc. These assays include CE as a complement to reverse-phase peptide mapping for the identification of small peptides eluting in the void volume, profiling N-linked glycopeptide heterogeneity, and determining O-linked site occupancy. In addition, CE-MS was used to confirm major 8-aminopyrene-1,3,6-trisulfonate (APTS)-labeled glycans released from recombinant antibodies that are routinely profiled by CE-laser-induced fluorescence (CE-LIF). For each study, CE-MS was able to successfully identify components seen in UV or LIF electropherograms, thereby expanding the capability of CE and CE-MS for profiling biomolecules.
Gennaro
Lynn A
LA
Late Stage Analytical Development, Genentech Inc., South San Francisco, CA 94080, USA. lgennaro@gene.com
Salas-Solano
Oscar
O
Ma
Stacey
S
D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D002240:Carbohydrate Sequence; D019075:Electrophoresis, Capillary / Q000379:methods*; D005453:Fluorescence; D006001:Glycoconjugates / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D007202:Indicators and Reagents; D007834:Lasers; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D010449:Peptide Mapping / Q000379:methods*; D011721:Pyrenes / Q000737:chemistry; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology / Q000378:metabolism
C103430:8-aminopyrene-1,3,6-trisulfonic acid; D000911:Antibodies, Monoclonal; D006001:Glycoconjugates; D007202:Indicators and Reagents; D011721:Pyrenes; D011994:Recombinant Proteins