Citation: WANG Yao, LIANG Gaodao, HAN Qing, HU Xun, ZHANG Qiwei, HE Zhenyu. Methylamidation and normal phase chromatography with silica-hydride-based stationary phase for N-glycosylation characterization of monoclonal antibody drugs[J]. Chinese Journal of Chromatography, ;2018, 36(7): 615-620. doi: 10.3724/SP.J.1123.2018.01026 shu

Methylamidation and normal phase chromatography with silica-hydride-based stationary phase for N-glycosylation characterization of monoclonal antibody drugs

WANG Yao ^1, ,
LIANG Gaodao ¹ ,
HAN Qing ¹ ,
HU Xun ¹ ,
ZHANG Qiwei ² ,
HE Zhenyu ¹

1.
Wuhan Centers for Disease Prevention & Control, Wuhan 430015, China;
2.
Jianghan University, Wuhan 430056, China

Received Date: 16 January 2018

Fund Project: General Program of Health and Family Planning Commission of Hubei Province(No. WJ2017M198).

Monoclonal antibodies (mAbs) are efficacious therapeutic agents against various complex diseases. N-glycosylation characterization of mAbs is based primarily on fluorescence derivatization coupled with hydrophilic interaction liquid chromatography (HILIC). However, this method has some drawbacks. Presently, the N-glycosylation of mAbs was analyzed by methylamidation and silica-hydride-based normal phase chromatography (SiH-NPC). Samples were analyzed by liquid chromatography-mass spectrometry after enzymatic cleavage, methylamidation, and purification. SiH-NPC had some advantages compared to HILIC. It offered a novel separation mechanism that allowed for high resolution using salt-free mobile phases, avoiding contamination of the mass spectrometer. SiH-NPC may be applicable for rapid analysis. Its structure was more stable than that of HILIC, and it provided a long service life. In combination with the sialic acid derivatization, SiH-NPC presented a significant advantage in the analysis of sialylated glycans as well as isomeric oligosaccharides by liquid chromatography-mass spectrometry. The approach has potential applications in the biopharmaceutical industry.
- methylamidation,
- silica-hydride,
- liquid chromatography-mass spectrometry(LC-MS),
- N-glycans,
- monoclonal antibodies(mAbs)
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