Abstract
Many proteins are glycosylated, that is to say they have bound sugars or glycans. Glycosylation is a non-template-driven posttranslation modification. It is now known that correct glycosylation is essential for the correct folding, solubility, stability, and immunogenicity of proteins. Here, we describe the technique of Lectin Affinity Chromatography (LAC), a procedure that has the ability to separate different glycans which are attached to proteins or lipids, termed glycoproteins or glycolipids, respectively. This method utilizes different immobilized lectins that have affinity for specific sugar substrates, to separate a wide range of glycan-attached complexes (Ambrosi et al., Org Biomol Chem 3:1593–1608, 2005). To further enhance the specificity of LAC, a corresponding free sugar may be used to produce a specific elution. In general, the conditions under which lectin affinity chromatography operates are relatively mild resulting in good biological recoveries of the glycoproteins.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Geyer H, Geyer R (2006) Strategies for analysis of glycoprotein glycosylation. Biochim Biophys Acta 1764:1853–1869
Qiu R, Regnier FE (2005) Use of multidimensional lectin affinity chromatography in differential glycoproteomics. Anal Chem 77:2802–2809
Wu A, Lisowska E, Duk M, Yang Z (2008) Lectins as tools in glycoconjugate research. Glycoconj J 26:899–913
Yang Z, Hancock WS (2005) Monitoring glycosylation pattern changes of glycoproteins using multi-lectin affinity chromatography. J Chromatogr A 1070:57–64
Ambrosi M, Cameron NR, Davis BG (2005) Lectins: tools for the molecular understanding of the glycocode. Org Biomol Chem 3:1593–1608
Lis H, Sharon N (1998) Lectins: carbohydrate-specific proteins that mediate cellular recognition. Chem Rev 98:637–674
Varki A, Etzler ME, Cummings RD, Esko JD (2009) Discovery and classification of glycanbinding proteins. In: Varki A, Cummings RD, Esko JD, Freeze HH, Stanley P, Bertozzi CR, Hart GW, Etzler ME (eds) Essentials of glycobiology, 2nd edn. Cold Spring Harbor Laboratory Press, New York, pp 375–386
Smith EA, Thomas WD, Kiessling LL, Corn RM (2003) Surface plasmon resonance imaging studies of protein-carbohydrate interactions. J Am Chem Soc 125:6140–6148
Mao X, Qin J, Lin B (2007) Miniaturized lectin affinity chromatography. In: Nilsson CL (ed) Lectin: analytical technologies. Elsevier B.V, Oxford, pp 213–238.
Keogh D, Thompson R, Larragy R, O’Connell M, O’Connor B, Clarke P (2014) Generating novel recombinant prokaryotic lectins with altered carbohydrate binding specificities and affinities through mutagenesis of the PA-IL protein from Pseudomonas aeruginosa. Biochim Biophys Acta 1840:2091–2104
Tateno HI, Nakamura-Tsuruta S, Hirabayashi J (2007) Frontal affinity chromatography: sugar-protein interactions. Nat Protoc 2:2529–2537
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media New York
About this protocol
Cite this protocol
O’Connor, B.F., Monaghan, D., Cawley, J. (2017). Lectin Affinity Chromatography (LAC). In: Walls, D., Loughran, S. (eds) Protein Chromatography. Methods in Molecular Biology, vol 1485. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6412-3_23
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6412-3_23
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6410-9
Online ISBN: 978-1-4939-6412-3
eBook Packages: Springer Protocols