Abstract
Glycosylation is a non-template-driven posttranslational modification during which linked-sugars and glycans are added to the nascent polypeptide. Over 70% of the eukaryotic proteome is thought to be glycosylated. It is now known that correct glycosylation is essential for the correct folding, solubility, stability, and immunogenicity of proteins. In this chapter, we describe the technique of lectin affinity chromatography (LAC), a procedure that has the ability to distinguish 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.
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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
Totten SM, Adusumilli R, Kullolli M, Tanimoto C, Brooks JD, Mallick P, Pitteri SJ (2018) Multi-lectin affinity chromatography and quantitative proteomic analysis reveal differential glycoform levels between prostate cancer and benign prostatic hyperplasia sera. Sci Rep 8(1):6509
Nauom S, da Silva Neto BR, Ribeiro MS, Pedersoli WR, Ulhoa CJ, Silva RN, Monteiro VN (2019) Biochemical and molecular study of Trichoderma harzianum enriched secretome protein profiles using lectin affinity chromatography. Appl Biochem Biotechnol 187(1):1–13
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, Cummings RD, Esko JD, Freeze HH, Stanley P, Bertozzi CR, Hart GW, Etzler ME (eds) (2009) Essentials of glycobiology. Cold Spring Harbor Laboratory Press, New York. isbn:978-087-969-770-9
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
Adamczyk B, Tharmalingam T, Rudd PM (2012) Glycans as cancer biomarkers. Biochim Biophys Acta 1820(9):1347–1353
Reily C, Stewart TJ, Renfrow MB et al (2019) Glycosylation in health and disease. Nat Rev Nephrol 15:346–366
Totten SM, Kullolli M, Pitteri SJ (2017) Multi-lectin affinity chromatography for separation, identification, and quantitation of intact protein glycoforms in complex biological mixtures. In: Proteomics. Humana Press, New York, pp 99–113
Hawkes N (2019) Cancer survival data emphasise importance of early diagnosis. BMJ 25(364):l408
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-proteininteractions. Nat Protoc 2:2529–2537
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O’Connor, B.F., Monaghan, D., Cawley, J. (2023). Lectin Affinity Chromatography. In: Loughran, S.T., Milne, J.J. (eds) Protein Chromatography. Methods in Molecular Biology, vol 2699. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3362-5_12
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DOI: https://doi.org/10.1007/978-1-0716-3362-5_12
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