title: Solid-phase reductive amination for glycomic analysis.
abstract: Reductive amination is an indispensable method for glycomic analysis, as it tremendously facilitates glycan characterization and quantification by coupling functional tags at the reducing ends of glycans. However, traditional in-solution derivatization based approach for the preparation of reductively aminated glycans is quite tedious and time-consuming. Here, a simpler and more efficient strategy termed solid-phase reductive amination was investigated. The general concept underlying this new approach is to streamline glycan extraction, derivatization, and purification on non-porous graphitized carbon sorbents. Neutral and sialylated standard glycans were utilized to test the feasibility of the solid-phase method. As results, almost complete labeling of those glycans with four common labels of aniline, 2-aminobenzamide (2-AB), 2-aminobenzoic acid (2-AA) and 2-amino-N-(2-aminoethyl)-benzamide (AEAB) was obtained, and negligible desialylation occurred during sample preparation. The labeled glycans derived from glycoproteins showed excellent reproducibility in high performance liquid chromatography (HPLC) and matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis. Direct comparisons based on fluorescent absorbance and relative quantification using isotopic labeling demonstrated that the solid-phase strategy enabled 20-30% increase in sample recovery. In short, the solid-phase strategy is simple, reproducible, efficient, and sensitive for glycan analysis. This method was also successfully applied for N-glycan profiling of HEK 293 cells with MALDI-TOF MS, showing its attractive application in the high-throughput analysis of mammalian glycome.
authors.affiliation: State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300353, China; Department of Chemistry, Georgia State University, Atlanta, GA 30303, United States.
authors.forename: Kuan
authors.initials: K
authors.lastname: Jiang
authors.affiliation: Department of Chemistry, Georgia State University, Atlanta, GA 30303, United States.
authors.forename: He
authors.initials: H
authors.lastname: Zhu
authors.affiliation: Department of Chemistry, Georgia State University, Atlanta, GA 30303, United States.
authors.forename: Cong
authors.initials: C
authors.lastname: Xiao
authors.affiliation: Department of Chemistry, Georgia State University, Atlanta, GA 30303, United States.
authors.forename: Ding
authors.initials: D
authors.lastname: Liu
authors.affiliation: Department of Chemistry, Georgia State University, Atlanta, GA 30303, United States.
authors.forename: Garrett
authors.initials: G
authors.lastname: Edmunds
authors.affiliation: Department of Chemistry, Georgia State University, Atlanta, GA 30303, United States.
authors.forename: Liuqing
authors.initials: L
authors.lastname: Wen
authors.affiliation: Department of Chemistry, Georgia State University, Atlanta, GA 30303, United States. Electronic address: cma@gsu.edu.
authors.forename: Cheng
authors.initials: C
authors.lastname: Ma
authors.affiliation: State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300353, China. Electronic address: Jinglink@nankai.edu.cn.
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.affiliation: State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300353, China; Department of Chemistry, Georgia State University, Atlanta, GA 30303, United States. Electronic address: pwang11@gsu.edu.
authors.forename: Peng George
authors.initials: PG
authors.lastname: Wang
mesh_terms: D000586:Amination; D054794:Glycomics / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D008745:Methylation; D010084:Oxidation-Reduction; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism
chemical_list: D011134:Polysaccharides
keywords: Glycomic analysis; Non-porous graphitized carbon; Reductive amination; Relative quantification; Solid-phase strategy