These authors contributed equally to this work.
Multiplex profiling of glycoproteins using a novel bead-based lectin array
Article first published online: 11 DEC 2013
© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Volume 14, Issue 1, pages 78–86, January 2014
How to Cite
Wang, H., Li, H., Zhang, W., Wei, L., Yu, H. and Yang, P. (2014), Multiplex profiling of glycoproteins using a novel bead-based lectin array. Proteomics, 14: 78–86. doi: 10.1002/pmic.201200544
Colour Online: See the article online to view Figs. 1 and 4 in colour
- Issue published online: 13 JAN 2014
- Article first published online: 11 DEC 2013
- Accepted manuscript online: 18 NOV 2013 07:09AM EST
- Manuscript Accepted: 23 OCT 2013
- Manuscript Revised: 17 OCT 2013
- Manuscript Received: 2 DEC 2012
- National Key Basic Research Program of China. Grant Numbers: 2011CB910600, 2012CB910602, 2013CB910502
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Figure S1. Signal curve made to obtain the optimum lectin amount per coupling reaction for (A) WGA, (B) AAL, (C) Con A, (D) PNA and (E) RCA120. The x-axis represents the various amounts of lectin coupled with 15 μL (approximately 1.875 × 105) beads. The y-axis represents the resulting fluorescence signal. The value of each spot on the y-axis presents the signal yield of each assay. The two gray curves represent negative controls. Error bars represent SD of triplicate determinants.
Figure S2. Competition assay of RCA120-ASF interaction with lactose. Error bars represent SD of triplicate determinants.
Figure S3. (A) LOD and (B) linearity of response of direct assay with Con A, AAL, PNA and WGA. (C) LOD of Hp through antibody-overlay profiling with SNA-I. Error bars represent SD of triplicate determinants.
Figure S4. The sensitivities of bead-based direct assay and antibody-overlay assay were compared and found to be basically the same. The limit of detection of lectin RCA120 was 0.5 ng/mL of human Hp in both array formats. Error bars represent SD of triplicate determinants.
Figure S5. (A) Multiplexed assay of RCA120 and PNA to test the specific interaction. ASF (1 μg/mL), RNB and BSA with same mole were used. (B) Dose-dependent net intensity of ASF (3 ng/mL to 1000 ng/mL) for RCA120 and PNA. Error bars represent SD of triplicate determinants.
Figure S6. (A) The normalized signals for RCA120, PNA, and Con A in single and multiplexed direct assay were statistically analyzed. 12 ng/mL of ASF was used. (B) The normalized signals for RCA120 and SNA-I in single and multiplexed antibody-overlay lectin array were statistically analyzed. 3 ng/mL of Hp was used. Error bars represent SD of triplicate determinants.
Table S1. Lectins used in this study.
Tabel S2. The optimum amounts per coupling reaction with five lectins (WGA, AAL, Con A, PNA, RCA120).
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