Identification of N-glycan of alpha-fetoprotein by lectin affinity microarray.
OBJECTIVE
The occurrence of cancer accompanies with changes in glycosylation of related proteins. A simple, rapid and high-throughput manner analyzing the N-glycan moiety is needed in the cancer glycoproteome study.


METHODS
Gel-slide was used as solid support of lectin microarray. We fabricated the lectin microarray with selected lectins to identify the N-glycan compositions of glycoproteins, to determine the individual N-glycan pattern of several glycoproteins and to compare the N-glycan compositions of alpha-fetoproteins (AFPs) from different sources.


RESULTS
It is feasible to analyze N-glycan pattern of glycoproteins with the lectin affinity microarray. The optimum loading concentration of lectins in this array is 1 mg/ml. The lectin microarray is sensitive, and it can even detect tested glycoprotein less than 1 pg. There is a linear relationship between the fluorescence intensity and the cy3 concentration of glycoprotein at the range from 10 to 1,000 nM.


CONCLUSIONS
The lectin microarray could give a panel of lectin affinity patterns of individual glycoproteins, and it could indicate the minimal differences of N-glycan compositions between AFPs from different sources. The developed lectin affinity microarray may contribute to the research of glycoproteomics associated with cancer and other diseases.
Chen
Pei
P
Liver Cancer Institute and Zhongshan Hospital, Fudan University, Shanghai, People's Republic of China.
Liu
YinKun
Y
Kang
XiaoNan
X
Sun
Lu
L
Yang
PengYuan
P
Tang
ZhaoYou
Z
D000818:Animals; D006023:Glycoproteins / Q000032:analysis / Q000737:chemistry; D006031:Glycosylation; D006801:Humans; D037102:Lectins / Q000737:chemistry*; D046228:Microarray Analysis / Q000379:methods*; D011134:Polysaccharides / Q000032:analysis*; D000509:alpha-Fetoproteins / Q000737:chemistry*
D006023:Glycoproteins; D037102:Lectins; D011134:Polysaccharides; D000509:alpha-Fetoproteins