Establishment of a method for determination of sialylation levels of N-glycan profiles based on ion exchange chromatography
10.19778/j.chp.2025.04.005
- VernacularTitle:基于离子交换色谱的高唾液酸N糖谱测定方法的建立
- Author:
Hongrui YIN
1
;
Xinxin FANG
1
;
Ying ZHANG
1
;
Mingming XU
1
;
Luxia ZHENG
1
;
Hong SHAO
1
Author Information
1. 上海市食品药品检验研究院,上海 201203
- Publication Type:Journal Article
- Keywords:
ion exchange chromatography;
sialylation levels of N-glycan profiles;
glycoprotein drugs
- From:
Drug Standards of China
2025;26(4):366-370
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a method for determining the sialylation levels of N-glycan profiles based on ion-exchange chromatography in glycoprotein drugs.Methods:The separation was performed in a column picking with anion-exchange resin(4.6 mm × 250 mm,5 μm,or equivalent).Mobile phase A was 20％ acetonitrile solution,while mobile phase B was a 20％acetonitrile solution containing 0.1 mol·L-1 ammonium formate(pH 4.5).The flow rate was 0.60 mL·min-1.Detection was performed using a fluorescence detector with excita-tion and emission wavelengths of 330 and 420 nm,respectively.The injection volume was 5 μL.Results:Within the target loading sample range of 40％ to 160％,the peak area showed a good linear relationship with protein concentration(r＞0.99).The average recovery rates at three concentration levels were 106.25％,100.00％,and 106.67％,respectively.The limit of quantification was 0.03％.Conclusion:This method is suitable for detecting the sialylation levels of N-glycan profiles in various glycoprotein drugs.
