Multiplex profiling of glycoproteins using a novel bead-based lectin array.
Lectin array is becoming important in profiling targeted glycan/glycoprotein, but weak interaction between lectin and glycan causes low sensitivity of the approach. This study aims to develop a bead-based lectin array for improving the sensitivity of glycosylation profiling. Lectins are chemically coupled to fluorescent dye coated microbeads, and glycan-lectin recognition is carried out three dimensionally. The performance of this platform was evaluated, and the LOD of lectin Ricinus communis agglutinin 120 (RCA120) was 50 pg/mL (1 pM) of asialofetuin, providing the bead-based lectin microarray with the highest sensitivity among the reported lectin microarrays. Furthermore, multiplexed assay was performed, which allowed the simultaneous detection of multiple carbohydrate epitopes in a single reaction vessel. The glycosylation patterns of hepatocellular carcinoma associated immunoglobulin G were analyzed, and increased (α-1,6) core fucosylation and (α-2,6) sialylation patterns were observed, which may provide significant clinical evidence for disease diagnosis.
Department of Chemistry, Fudan University, Shanghai, China; Institutes of Biomedical Sciences, Fudan University, Shanghai, China; Shanghai Institute for food and drug control, Shanghai, China.
Hong
H
Wang
Hong
H
Li
Wei
W
Zhang
Liming
L
Wei
Hongxiu
H
Yu
Pengyuan
P
Yang
D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood / Q000737:chemistry; D037102:Lectins / Q000737:chemistry* / Q000378:metabolism; D057230:Limit of Detection; D008113:Liver Neoplasms / Q000097:blood; D008863:Microspheres; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results
D006023:Glycoproteins; D007074:Immunoglobulin G; D037102:Lectins
Bead-based lectin array; Glycan profiling; Glycoproteomics; Multiplexed assay; Sensitivity