Online Hydrophobic Interaction Chromatography-Mass Spectrometry for the Analysis of Intact Monoclonal Antibodies

被引：60

作者：

Chen, Bifan ^{[1
]}

Lin, Ziqing ^{[2
,3
]}

Alpert, Andrew J. ^{[4
]}

Fu, Cexiong ^{[5
]}

Zhang, Qunying ^{[5
]}

Pritts, Wayne A. ^{[5
]}

Ge, Ying ^{[1
,2
,3
]}

机构：

[1] Univ Wisconsin, Dept Chem, Madison, WI 53705 USA

[2] Univ Wisconsin, Dept Cell & Regenerat Biol, Madison, WI 53705 USA

[3] Univ Wisconsin, Sch Med & Publ Hlth, Human Prote Program, Madison, WI 53705 USA

[4] PolyLC Inc, Columbia, MD 21045 USA

[5] AbbVie Inc, Proc Analyt, N Chicago, IL 60064 USA

来源：

ANALYTICAL CHEMISTRY | 2018年 / 90卷 / 12期

关键词：

TOP-DOWN PROTEOMICS; THERAPEUTIC ANTIBODIES; BIOPHYSICAL CHARACTERIZATION; SEPARATION; FUTURE;

D O I：

10.1021/acs.analchem.8b01865

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Therapeutic monoclonal antibodies (mAbs) are an important class of drugs for a wide spectrum of human diseases. Liquid chromatography (LC) coupled to mass spectrometry (MS) is one of the techniques in the forefront for comprehensive characterization of analytical attributes of mAbs. Among various protein chromatography modes, hydrophobic interaction chromatography (HIC) is a popular offline nondenaturing separation technique utilized to purify and analyze mAbs, typically with the use of non-MS-compatible mobile phases. Herein we demonstrate for the first time, the application of direct HIC-MS and HIC-tandem MS (MS/MS) with electron capture dissociation (ECD) for analyzing intact mAbs on quadrupole-time-of-flight (Q-TOF) and Fourier transform ion cyclotron resonance (FTICR) mass spectrometers, respectively. determination of relative hydrophobicity, intact masses, and glycosylation profiles of mAbs as well as sequence and structural characterization of the complementarity-determining regions in an online configuration.

引用

页码：7135 / 7138

页数：4

共 25 条

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