Lectin Microarrays
Our lab developed lectin microarrays for rapid analysis of the glycome (1-3). These microarrays utilize immobilized carbohydrate-binding proteins at high spatial density to give specific information on the repertoire of glycans present, e.g., high mannose epitopes, branching patterns, and terminal α-2,3- or α-2,6-sialic acids, in a high-throughput format. We directly mimic how nature recognizes glycans with the use of lectin probes, i.e., via 3-5 glycan binding epitopes, and gain discrete structural information on glycan linkages.
Quick Reference
Benefits of Using Lectin Microarrays
Unlike other glycomic methodologies, lectin microarrays:
- Can directly observe the N-linked, O-linked and glycolipid glycomes simultaneously (2)
- Require minimal manipulation or derivatization of samples, allowing low amounts of raw starting material to be analyzed (100 μg or less of raw material is routinely used in our lab, compared to 3-5 mg for a typical glycomic mass spectrometry analysis) and
- Generate data in the same format as mRNA and miRNA datasets, enabling facile data integration.
Our unique dual-color methodology allows for more accurate semi-quantitative analysis of the glycome. We print our own lectin microarrays, which include >90 plant lectins, recombinant lectins, and selected antibodies (4, 5). The discrete specificities of our probes are well characterized and many of them have been analyzed using the Consortium for Functional Glycomics glycan microarray (6).
Annotations of Lectin Binding Specificities
Glycomic Technology: Timeline of Development
Lectin microarray technology for the rapid, simple survey of protein glycosylation (1)
Two-color lectin microarray approach to evaluate differences in the glycosylation of heterogeneous samples (2)
Strategy to systematically create a recombinant, well-defined lectin set for use in microarray technology (4)
Oriented lectin microarray platform to improve overall sensitivity and limit detection of recombinant lectins by site-specific orientation (7)
Expression of bacterially-derived lectins and their application to a recombinant lectin microarray (8)
One-pot method for the fabrication of a localized GSH-scaffold to orient GST-tagged lectins in situ (9)
Updated lectin microarray protocol that uses a non-contact, piezoelectric printer for increased lectin activity on the array (3)
Integration of microarray data with 3D structural modeling (10)
Use of machine learning for large-scale evaluation of binding motifs in glycan array data (11)
- Pilobello, K.T.; Krishnamoorthy, L.; Slawek, D.; Mahal, L.K. Development of a lectin microarray for the rapid analysis of protein glycopatterns. ChemBioChem, 2005, 6, 985-989. doi: 10.1002/cbic.200400403
- Pilobello, K.T.; Slawek, D.; Mahal, L.K. A ratiometric lectin microarray approach to analysis of the dynamic mammalian glycome, Proc. Natl. Acad. Sci., USA, 2007, 104, 10534-10539. doi: 10.1073/pnas.0704954104
- Pilobello, K.T; Agrawal, P.; Rouse, R.; Mahal, L.K. Advances in lectin microarray technology: Optimized protocols for piezoelectric print conditions. Curr. Prot. Chem. Biol., 2013, 5, 1-23. doi: 10.1002/9780470559277.ch120035
- Hsu, K.-L.; Gildersleeve, J.C.; Mahal, L.K. A simple strategy for the creation of a recombinant lectin microarray, Mol. BioSystems, 2008, 4, 654-662. doi: 10.1039/b800725j
- Ribeiro, J.P.; Pau, W.K.; Pifferi, C.; Renaudet, O; Varrot, A; Mahal, L.K. ‡; Imberty, A. ‡ Characterization of a high-affinity sialic acid specific CBM40 from Clostridium perfringens and engineering of a divalent form. Biochem. J., 2016, 473, 2109-18. doi: 10.1042/BCJ20160340. ‡ Co-corresponding authors.
- Wang, L.; Cummings, R.D.; Smith, D.F.; Huflejt, M.; Campbell, C.T.; Gildersleeve, J.D.; Gerlach, J.Q.; Kilcoyne, M.; Joshi, L.; Serna, S.; Reichardt, N.-C.; Pera, N.P.; Pieters, R.; Eng, W.S.; Mahal, L.K. Cross-platform comparison of glycan microarray formats. Glycobiology, 2014, 24, 507-517. doi: 10.1093/glycob/cwu019
- Propheter, D.C.; Hsu, K.-L.; Mahal, L.K. Fabrication of an oriented lectin microarray, ChemBioChem, 2010, 11, 1203-1207. doi: 10.1002/cbic.201000106
- Propheter, D.C.; Hsu, K.-L.; Mahal, L.K. Recombinant lectin microarrays for glycomic analysis. Methods Mol. Biol., 2011, 723, 67-77. doi: 10.1007/978-1-61779-043-0_6
- Propheter, D.C.; Mahal, L.K. Orientation of GST-tagged lectins via in situ surface modification to create an expanded lectin microarray for glycomic analysis. Mol. Biosystems, 2011, 7, 2114-7. doi: 10.1039/c1mb05047h
- Grant, O.C.; Tessier, M.B.; Meche, L.; Mahal, L.K.; Foley, B.L.; Woods, R.J. Combining 3D structure with glycan array data provides insight into the origin of glycan specificity. Glycobiology, 2016, 26, 772-83. doi: 10.1093/glycob/cww020
- Bojar, D.; Meche, L.; Meng, G.; Eng, W.; Smith, D.F.; Cummings, R.D.; Mahal, L.K. A useful guide to lectin binding: Machine-learning directed annotation of 57 unique lectin specificities. ACS Chemical Biology, 2022, in Press. Formerly: bioRixv, doi: 10.1101/2021.08.31.458439
Get in Touch
- mahaladm@ualberta.ca
- (780) 492-5847
- W3-39B Chemistry Centre, 11227 Saskatchewan Drive NW, Edmonton, AB T6G 2G2
F.A.Q.
The Canada Excellence Research Chairs (CERC) Program awards universities up to $10 million over seven years to support world‑renowned researchers and their teams to establish ambitious research programs at Canadian universities.
Located in beautiful Edmonton, Alberta, Canada, the Chemistry Department at the UofA is renown for its active research. It is one of the best equipped and well funded departments in Canada.
It is a glycomic technology developed in the Mahal Lab, provide a rapid analysis of the glycome (1-3). These microarrays utilize immobilized carbohydrate-binding proteins at high spatial density to give specific information on the repertoire of glycans present.
miRNA can be used to identify glycosylation enzymes and their corresponding glycans that drive disease states.
At the UofA, you do not have to find a supervisor in order to apply. The department has a process in place for students to find a supervisor and join a research group once they arrive to start their program. However, it is recommended for applicants to contact chemistry faculty members whose areas of research are of interest to you. Information about requirements and how to apply can be found here.