A rapid and simple approach for glycoform analysis.
Fast glycoform analysis is important for quality control of glycoproteins that account for over 40% of the approved biopharmaceuticals. Herein, we realized an Au nanoparticle-based lectin affinity chromatography (LAC) using simple standard laboratory equipment for fast glycoform analysis. Pisum sativum agglutinin (PA), a lectin derived from P. sativum, was covalently conjugated to Au nanoparticles via naturally formed carboxylic groups onto the surface of Au nanoparticles and amino groups of PA. Each model glycoprotein was separated into several fractions including the unbound, weakly bound, modestly bound, and strongly bound glycoforms based on affinity strength of the glycoform toward PA. A single run of Au nanoparticle-based LAC was finished within 18 min, which could be further decreased by centrifuging the mixture of the PA functionalized Au nanoparticles and the glycoproteins at a higher speed. To our knowledge, we are the first to use Au nanoparticles as LAC matrix.
School of Chemistry and Chemical Engineering, Jiangsu Key Laboratory of Green Synthesis for Functional Material, Jiangsu Normal University, Xuzhou, Jiangsu 221116, China.
Xiaojun
X
Liu
School of Chemistry and Chemical Engineering, Jiangsu Key Laboratory of Green Synthesis for Functional Material, Jiangsu Normal University, Xuzhou, Jiangsu 221116, China.
Yawei
Y
Wang
School of Chemistry and Chemical Engineering, Jiangsu Key Laboratory of Green Synthesis for Functional Material, Jiangsu Normal University, Xuzhou, Jiangsu 221116, China.
Yang
Y
Tu
School of Chemistry and Chemical Engineering, Jiangsu Key Laboratory of Green Synthesis for Functional Material, Jiangsu Normal University, Xuzhou, Jiangsu 221116, China.
Ziqi
Z
Zhu
School of Chemistry and Chemical Engineering, Jiangsu Key Laboratory of Green Synthesis for Functional Material, Jiangsu Normal University, Xuzhou, Jiangsu 221116, China.
Xiaogang
X
Li
School of Chemistry and Chemical Engineering, Jiangsu Key Laboratory of Green Synthesis for Functional Material, Jiangsu Normal University, Xuzhou, Jiangsu 221116, China.
Qingquan
Q
Zhang
School of Chemistry and Chemical Engineering, Jiangsu Key Laboratory of Green Synthesis for Functional Material, Jiangsu Normal University, Xuzhou, Jiangsu 221116, China.
Wenfeng
W
Zhao
School of Chemistry and Chemical Engineering, Jiangsu Key Laboratory of Green Synthesis for Functional Material, Jiangsu Normal University, Xuzhou, Jiangsu 221116, China.
Yuling
Y
Li
School of Chemistry and Chemical Engineering, Jiangsu Key Laboratory of Green Synthesis for Functional Material, Jiangsu Normal University, Xuzhou, Jiangsu 221116, China. Electronic address: gai@jsnu.edu.cn.
Hongwei
H
Gai
D000373:Agglutinins / Q000737:chemistry; D002846:Chromatography, Affinity; D006023:Glycoproteins / Q000032:analysis*; D006046:Gold / Q000737:chemistry; D037102:Lectins / Q000737:chemistry; D053768:Metal Nanoparticles / Q000737:chemistry; D018532:Pisum sativum
D000373:Agglutinins; D006023:Glycoproteins; D037102:Lectins; D006046:Gold
Au nanoparticles; Glycoform analysis; Lectin affinity chromatography; Quality control