PURPOSE: 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. CONCLUSION: 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.
PURPOSE: 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. CONCLUSION: 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.
Authors: One Choi; Noboru Tomiya; Jung H Kim; James M Slavicek; Michael J Betenbaugh; Yuan C Lee Journal: Glycobiology Date: 2003-04-02 Impact factor: 4.313
Authors: Leonardo Nimrichter; Ari Gargir; Monica Gortler; Rom T Altstock; Avi Shtevi; Oori Weisshaus; Ella Fire; Nir Dotan; Ronald L Schnaar Journal: Glycobiology Date: 2003-11-24 Impact factor: 4.313
Authors: William R Alley; Jacqueline A Vasseur; John A Goetz; Martin Svoboda; Benjamin F Mann; Daniela E Matei; Nancy Menning; Ahmed Hussein; Yehia Mechref; Milos V Novotny Journal: J Proteome Res Date: 2012-03-07 Impact factor: 4.466
Authors: Chen Li; Diane M Simeone; Dean E Brenner; Michelle A Anderson; Kerby A Shedden; Mack T Ruffin; David M Lubman Journal: J Proteome Res Date: 2009-02 Impact factor: 4.466
Authors: Jared Q Gerlach; Anja Krüger; Susan Gallogly; Shirley A Hanley; Marie C Hogan; Christopher J Ward; Lokesh Joshi; Matthew D Griffin Journal: PLoS One Date: 2013-09-19 Impact factor: 3.240