Capillary electrophoresis-mass spectrometry as a characterization tool for therapeutic proteins

被引：62

作者：

Gennaro, Lynn A. ^{[1
]}

Salas-Solano, Oscar ^{[1
]}

Ma, Stacey ^{[1
]}

机构：

[1] Genentech Inc, Late Stage Analyt Dev, San Francisco, CA 94080 USA

来源：

ANALYTICAL BIOCHEMISTRY | 2006年 / 355卷 / 02期

关键词：

capillary electrophoresis; mass spectrometry; peptide digests; glycoproteins; carbohydrates; APTS;

D O I：

10.1016/j.ab.2006.04.002

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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. (c) 2006 Elsevier Inc. All rights reserved.

引用

页码：249 / 258

页数：10

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