Glycoconjugate Journal

, Volume 30, Issue 2, pp 137–146 | Cite as

Strategies for the profiling, characterisation and detailed structural analysis of N-linked oligosaccharides

  • Tharmala Tharmalingam
  • Barbara Adamczyk
  • Margaret A. Doherty
  • Louise Royle
  • Pauline M. Rudd


Many post-translational modifications, including glycosylation, are pivotal for the structural integrity, location and functional activity of glycoproteins. Sub-populations of proteins that are relocated or functionally changed by such modifications can change resting proteins into active ones, mediating specific effector functions, as in the case of monoclonal antibodies. To ensure safe and efficacious drugs it is essential to employ appropriate robust, quantitative analytical strategies that can (i) perform detailed glycan structural analysis, (ii) characterise specific subsets of glycans to assess known critical features of therapeutic activities (iii) rapidly profile glycan pools for at-line monitoring or high level batch to batch screening. Here we focus on these aspects of glycan analysis, showing how state-of-the-art technologies are required at all stages during the production of recombinant glycotherapeutics. These data can provide insights into processing pathways and suggest markers for intervention at critical control points in bioprocessing and also critical decision points in disease and drug monitoring in patients. Importantly, these tools are now enabling the first glycome/genome studies in large populations, allowing the integration of glycomics into other ‘omics platforms in a systems biology context.


Glycosylation Glycan analysis High throughput Glycoprofiling Database 





Antibody dependent cell cytotoxicity




Ethylene Bridged Hybrid


Capillary electrophoresis with laser induced fluorescence


Critical quality attribute


European Medicines Agency




Electrospray ionization – mass spectrometry


Food and Drug Administration


Fluorescence detector

FUT6 and FUT8

Fucosyltransferase 6 and 8


Genome-wide association study


Hydrophilic interaction chromatography


Hepatocyte nuclear factor 1α


High performance liquid chromatography


Immunoglobulin G


Liquid chromatography – mass spectrometry


Matrix-assisted laser desorption/ionization


Nuclear magnetic resonance


Process analytical technology


Porous graphitized carbon


Reversed phase


Single nucleotide polymorphisms


Ultra performance liquid chromatography


Weak anion exchange



TT acknowledges funding from Science Foundation Ireland (Reproductive Biology Research Cluster (RBRC)) [grant number 07/SRC/B1156], BA acknowledges funding from the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement 215536 (EuroGlycoArrays), MAD acknowledges funding from the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement 259869 (GlycoBioM). The authors thank Prof. Johannis P. Kamerling for fruitful discussions.


  1. 1.
    Shahrokh, Z., Royle, L., Saldova, R., Bones, J., Abrahams, J.L., Artemenko, N.V., Flatman, S., Davies, M., Baycroft, A., Sehgal, S., Heartlein, M.W., Harvey, D.J., Rudd, P.M.: Erythropoietin produced in a human cell line (Dynepo) has significant differences in glycosylation compared with erythropoietins produced in CHO cell lines. Mol. Pharm. 8(1), 286–296 (2010). doi:10.1021/mp100353a PubMedCrossRefGoogle Scholar
  2. 2.
    Marino, K., Bones, J., Kattla, J.J., Rudd, P.M.: A systematic approach to protein glycosylation analysis: a path through the maze. Nat. Chem. Biol. 6(10), 713–723 (2010)PubMedCrossRefGoogle Scholar
  3. 3.
    Campbell, M.P., Royle, L., Radcliffe, C.M., Dwek, R.A., Rudd, P.M.: GlycoBase and autoGU: tools for HPLC-based glycan analysis. Bioinformatics 24(9), 1214–1216 (2008). doi:10.1093/bioinformatics/btn090 PubMedCrossRefGoogle Scholar
  4. 4.
    Hilliard, M., Struwe, W., Carta, G., O’Rourke, J., McLoughlin, N., Rudd, P.M., Qing, Y.: A systematic approach to glycan analysis using HILIC-UPLC and an online database of standardized values. In: (2012).
  5. 5.
    Guillarme, D., Ruta, J., Rudaz, S., Veuthey, J.L.: New trends in fast and high-resolution liquid chromatography: a critical comparison of existing approaches. Anal. Bioanal. Chem. 397(3), 1069–1082 (2010). doi:10.1007/s00216-009-3305-8 PubMedCrossRefGoogle Scholar
  6. 6.
    Novakova, L., Matysova, L., Solich, P.: Advantages of application of UPLC in pharmaceutical analysis. Talanta 68(3), 908–918 (2006). doi:10.1016/j.talanta.2005.06.035 PubMedCrossRefGoogle Scholar
  7. 7.
    Lame, M.E., Chambers, E.E., Blatnik, M.: Quantitation of amyloid beta peptides Abeta(1–38), Abeta(1–40), and Abeta(1–42) in human cerebrospinal fluid by ultra-performance liquid chromatography-tandem mass spectrometry. Anal. Biochem. 419(2), 133–139 (2011). doi:10.1016/j.ab.2011.08.010 PubMedCrossRefGoogle Scholar
  8. 8.
    Guile, G.R., Rudd, P.M., Wing, D.R., Prime, S.B., Dwek, R.A.: A rapid high-resolution high-performance liquid chromatographic method for separating glycan mixtures and analyzing oligosaccharide profiles. Anal. Biochem. 240(2), 210–226 (1996)PubMedCrossRefGoogle Scholar
  9. 9.
    Royle, L., Campbell, M.P., Radcliffe, C.M., White, D.M., Harvey, D.J., Abrahams, J.L., Kim, Y.-G., Henry, G.W., Shadick, N.A., Weinblatt, M.E., Lee, D.M., Rudd, P.M., Dwek, R.A.: HPLC-based analysis of serum N-glycans on a 96-well plate platform with dedicated database software. Anal. Biochem. 376(1), 1–12 (2008)PubMedCrossRefGoogle Scholar
  10. 10.
    Ahn, J., Bones, J., Yu, Y.Q., Rudd, P.M., Gilar, M.: Separation of 2-aminobenzamide labeled glycans using hydrophilic interaction chromatography columns packed with 1.7um sorbent. J Chromatogr B 878(3:4), 403–408 (2010)CrossRefGoogle Scholar
  11. 11.
    Pucic, M., Knezevic, A., Vidic, J., Adamczyk, B., Novokmet, M., Polasek, O., Gornik, O., Supraha-Goreta, S., Wormald, M.R., Redzic, I., Campbell, H., Wright, A., Hastie, N.D., Wilson, J.F., Rudan, I., Wuhrer, M., Rudd, P.M., Josic, D., Lauc, G.: High throughput isolation and glycosylation analysis of igg–Variability and heritability of the IgG glycome in three isolated human populations”. Mol. Cell Proteomics 10(10), (2011). doi:10.1074/mcp.M111.010090
  12. 12.
    Wormald, M.R., Rudd, P.M., Harvey, D.J., Chang, S.C., Scragg, I.G., Dwek, R.A.: Variations in oligosaccharide‐protein interactions in immunoglobulin G determine the site‐specific glycosylation profiles and modulate the dynamic moon of the Fc oligosaccharides. Biochemistry 36(6), 1370–1380 (1997)Google Scholar
  13. 13.
    Bones, J., McLoughlin, N., Hilliard, M., Wynne, K., Karger, B.L., Rudd, P.M.: 2D-LC analysis of BRP 3 erythropoietin N-glycosylation using anion exchange fractionation and hydrophilic interaction UPLC reveals long poly-N-acetyl lactosamine extensions. Anal. Chem. 83(11), 4154–4162 (2011). doi:10.1021/ac200406z PubMedCrossRefGoogle Scholar
  14. 14.
    Bones, J., Mittermayr, S., O’Donoghue, N., Guttman, A., Rudd, P.M.: Ultra performance liquid chromatographic profiling of serum N-glycans for fast and efficient identification of cancer associated alterations in glycosylation. Anal. Chem. 82(24), 10208–10215 (2010). doi:10.1021/ac102860w PubMedCrossRefGoogle Scholar
  15. 15.
    Huffman, J.E., Knezevic, A., Vitart, V., Kattla, J., Adamczyk, B., Novokmet, M., Igl, W., Pucic, M., Zgaga, L., Johannson, A., Redzic, I., Gornik, O., Zemunik, T., Polasek, O., Kolcic, I., Pehlic, M., Koeleman, C.A.M., Campbell, S., Wild, S.H., Hastie, N.D., Campbell, H., Gyllensten, U., Wuhrer, M., Wilson, J.F., Hayward, C., Rudan, I., Rudd, P.M., Wright, A.F., Lauc, G.: Polymorphisms in B3GAT1, SLC9A9 and MGAT5 are associated with variation within the human plasma N-glycome of 3533 European adults. Hum. Mol. Genet. 20(24), 5000–5011 (2011). doi:10.1093/hmg/ddr414 PubMedCrossRefGoogle Scholar
  16. 16.
    Domann, P.J., Pardos-Pardos, A.C., Fernandes, D.L., Spencer, D.I.R., Radcliffe, C.M., Royle, L., Dwek, R.A., Rudd, P.M.: Separation-based glycoprofiling approaches using fluorescent labels. Proteomics 7(S1), 70–76 (2007). doi:10.1002/pmic.200700640 PubMedCrossRefGoogle Scholar
  17. 17.
    Szabo, Z., Guttman, A., Karger, B.L.: Rapid release of N-linked glycans from glycoproteins by pressure-cycling technology. Anal. Chem. 82(6), 2588–2593 (2010). doi:10.1021/ac100098e PubMedCrossRefGoogle Scholar
  18. 18.
    Olajos, M., Hajos, P., Bonn, G.K., Guttman, A.: Sample preparation for the analysis of complex carbohydrates by multicapillary gel electrophoresis with light-emitting diode induced fluorescence detection. Anal. Chem. 80(11), 4241–4246 (2008). doi:10.1021/ac8002598 PubMedCrossRefGoogle Scholar
  19. 19.
    Szabo, Z., Guttman, A., Rejtar, T., Karger, B.L.: Improved sample preparation method for glycan analysis of glycoproteins by CE-LIF and CE-MS. Electrophoresis 31(8), 1389–1395 (2010). doi:10.1002/elps.201000037 PubMedCrossRefGoogle Scholar
  20. 20.
    Ruhaak, L.R., Hennig, R., Huhn, C., Borowiak, M., Dolhain, R.J., Deelder, A.M., Rapp, E., Wuhrer, M.: Optimized workflow for preparation of APTS-labeled N-glycans allowing high-throughput analysis of human plasma glycomes using 48-channel multiplexed CGE-LIF. J. Proteome Res. 9(12), 6655–6664 (2010). doi:10.1021/pr100802f PubMedCrossRefGoogle Scholar
  21. 21.
    Mittermayr, S., Bones, J., Doherty, M., Guttman, A., Rudd, P.M.: Multiplexed analytical glycomics: rapid and confident IgG N-glycan structural elucidation. J. Proteome Res. 10(8), 3820–3829 (2011). doi:10.1021/pr200371s PubMedCrossRefGoogle Scholar
  22. 22.
    Vanderschaeghe, D., Szekrenyes, A., Wenz, C., Gassmann, M., Naik, N., Bynum, M., Yin, H., Delanghe, J., Guttman, A., Callewaert, N.: High-throughput profiling of the serum N-glycome on capillary electrophoresis microfluidics systems: toward clinical implementation of GlycoHepatoTest. Anal. Chem. 82(17), 7408–7415 (2010). doi:10.1021/ac101560a PubMedCrossRefGoogle Scholar
  23. 23.
    North, S.J., Huang, H.-H., Sundaram, S., Jang-Lee, J., Etienne, A.T., Trollope, A., Al-Chalabi, S., Dell, A., Stanley, P., Haslam, S.M.: Glycomics profiling of Chinese hamster ovary (CHO) cell glycosylation mutants reveals N-glycans of a novel size and complexity. J. Biol. Chem. (2009). doi:10.1074/jbc.M109.068353
  24. 24.
    Hua, S., Lebrilla, C., An, H.J.: Application of nano-LC-based glycomics towards biomarker discovery. Bioanalysis 3(22), 2573–2585 (2011). doi:10.4155/bio.11.263 PubMedCrossRefGoogle Scholar
  25. 25.
    Kirsch, S., Bindila, L.: Nano-LC and HPLC-chip-ESI-MS: an emerging technique for glycobioanalysis. Bioanalysis 1(7), 1307–1327 (2009). doi:10.4155/bio.09.110 PubMedCrossRefGoogle Scholar
  26. 26.
    Kosicek, M., Kirsch, S., Bene, R., Trkanjec, Z., Titlic, M., Bindila, L., Peter-Katalinic, J., Hecimovic, S.: Nano-HPLC-MS analysis of phospholipids in cerebrospinal fluid of Alzheimer’s disease patients–a pilot study. Anal. Bioanal. Chem. 398(7–8), 2929–2937 (2010). doi:10.1007/s00216-010-4273-8 PubMedCrossRefGoogle Scholar
  27. 27.
    Ruhaak, L.R., Deelder, A.M., Wuhrer, M.: Oligosaccharide analysis by graphitized carbon liquid chromatography-mass spectrometry. Anal. Bioanal. Chem. 394(1), 163–174 (2009). doi:10.1007/s00216-009-2664-5 PubMedCrossRefGoogle Scholar
  28. 28.
    Wohlgemuth, J., Karas, M., Jiang, W., Hendriks, R., Andrecht, S.: Enhanced glyco-profiling by specific glycopeptide enrichment and complementary monolithic nano-LC (ZIC-HILIC/RP18e)/ESI-MS analysis. J. Sep. Sci. 33(6–7), 880–890 (2010). doi:10.1002/jssc.200900771 PubMedCrossRefGoogle Scholar
  29. 29.
    Zamfir, A., Vakhrushev, S., Sterling, A., Niebel, H.J., Allen, M., Peter-Katalinic, J.: Fully automated chip-based mass spectrometry for complex carbohydrate system analysis. Anal. Chem. 76(7), 2046–2054 (2004). doi:10.1021/ac035320q PubMedCrossRefGoogle Scholar
  30. 30.
    Zamfir, A.D., Bindila, L., Lion, N., Allen, M., Girault, H.H., Peter-Katalinic, J.: Chip electrospray mass spectrometry for carbohydrate analysis. Electrophoresis 26(19), 3650–3673 (2005). doi:10.1002/elps.200500101 PubMedCrossRefGoogle Scholar
  31. 31.
    Hua, S., An, H.J., Ozcan, S., Ro, G.S., Soares, S., DeVere-White, R., Lebrilla, C.B.: Comprehensive native glycan profiling with isomer separation and quantitation for the discovery of cancer biomarkers. Analyst 136(18), 3663–3671 (2011). doi:10.1039/c1an15093f PubMedCrossRefGoogle Scholar
  32. 32.
    Bynum, M.A., Yin, H., Felts, K., Lee, Y.M., Monell, C.R., Killeen, K.: Characterization of IgG N-glycans employing a microfluidic chip that integrates glycan cleavage, sample purification, LC separation, and MS detection. Anal. Chem. 81(21), 8818–8825 (2009). doi:10.1021/ac901326u PubMedCrossRefGoogle Scholar
  33. 33.
    Yu, Y.Q.: Analysis of N-linked glycans from coagulation factor IX, recombinant and plasma derived, using HILIC UPLC/FLR/QTof MS. In: Waters (ed.) Application Note, (2011).
  34. 34.
    Yu, Y.Q., Ahn, J., Gilar, M.: Trastuzumab glycan batch-to-batch profiling using a UPLC/FLR/MS platform In: Waters (ed.) Application Note, (2011).
  35. 35.
    Kattla, J.J., Struwe, W.B., Campbell, M.P., Doherty, M., Adamczyk, B., Saldova, R., Rudd, P.M.: Protein glycosylation. In: Moo-Young, M. (ed.) Comprehensive Biotechnology, vol. 3. pp. 467–486. Elsevier (2011).
  36. 36.
    Fulton, S., Murphy, S., Reich, J., Van Den Heuvel, Z., Sakowski, R., Smith, R., Agee, S.: A high-throughput microchromatography platform for quantitative analytical scale protein sample preparation. J. Assoc. Lab. Autom. 16(6), 457–467 (2011)CrossRefGoogle Scholar
  37. 37.
    von der Lieth, C.-W., Freire, A.A., Blank, D., Campbell, M.P., Ceroni, A., Damerell, D.R., Dell, A., Dwek, R.A., Ernst, B., Fogh, R., Frank, M., Geyer, H., Geyer, R., Harrison, M.J., Henrick, K., Herget, S., Hull, W.E., Ionides, J., Joshi, H.J., Kamerling, J.P., Leeflang, B.R., Lutteke, T., Lundborg, M., Maass, K., Merry, A., Ranzinger, R., Rosen, J., Royle, L., Rudd, P.M., Schloissnig, S., Stenutz, R., Vranken, W.F., Widmalm, G., Haslam, S.M.: EUROCarbDB: an open-access platform for glycoinformatics. Glycobiology 21(4), 493–502 (2011). doi:10.1093/glycob/cwq188 PubMedCrossRefGoogle Scholar
  38. 38.
    Ceroni, A., Maass, K., Geyer, H., Geyer, R., Dell, A., Haslam, S.M.: GlycoWorkbench: a tool for the computer-assisted annotation of mass spectra of glycans. J. Proteome Res. 7(4), 1650–1659 (2008). doi:10.1021/pr7008252 PubMedCrossRefGoogle Scholar
  39. 39.
    Maass, K., Ranzinger, R., Geyer, H., von der Lieth, C.-W., Geyer, R.: “Glyco-peakfinder” – de novo composition analysis of glycoconjugates. Proteomics 7(24), 4435–4444 (2007). doi:10.1002/pmic.200700253 PubMedCrossRefGoogle Scholar
  40. 40.
    Jansson, P.-E., Stenutz, R., Widmalm, G.R.: Sequence determination of oligosaccharides and regular polysaccharides using NMR spectroscopy and a novel Web-based version of the computer program casper. Carbohydr. Res. 341(8), 1003–1010 (2006)PubMedCrossRefGoogle Scholar
  41. 41.
    Campbell, M.P., Hayes, C.A., Struwe, W.B., Wilkins, M.R., Aoki-Kinoshita, K.F., Harvey, D.J., Rudd, P.M., Kolarich, D., Lisacek, F., Karlsson, N.G., Packer, N.H.: UniCarbKB: putting the pieces together for glycomics research. Proteomics 11(21), 4117–4121 (2011). doi:10.1002/pmic.201100302 PubMedCrossRefGoogle Scholar
  42. 42.
    Adamczyk, B., Tharmalingam, T., Rudd, P.M.: Glycans as cancer biomarkers. Biochim. Biophys. Acta (BBA) Gen. Subj. (0), (2012). Accessed 9 Dec 2011
  43. 43.
    Ercan, A., Cui, J., Chatterton, D.E.W., Deane, K.D., Hazen, M.M., Brintnell, W., O’Donnell, C.I., Derber, L.A., Weinblatt, M.E., Shadick, N.A., Bell, D.A., Cairns, E., Solomon, D.H., Holers, V.M., Rudd, P.M., Lee, D.M.: Aberrant IgG galactosylation precedes disease onset, correlates with disease activity, and is prevalent in autoantibodies in rheumatoid arthritis. Arthritis Rheum. 62(8), 2239–2248 (2010). doi:10.1002/art.27533 PubMedCrossRefGoogle Scholar
  44. 44.
    Coman, D.J., Murray, D.W., Byrne, J.C., Rudd, P.M., Bagaglia, P.M., Doran, P.D., Treacy, E.P.: Galactosemia, a single gene disorder with epigenetic consequences. Pediatr. Res. 67(3), 286–292 (2010)PubMedCrossRefGoogle Scholar
  45. 45.
    Coss, K.P., Byrne, J.C., Coman, D.J., Adamczyk, B., Abrahams, J.L., Saldova, R., Brown, A.Y., Walsh, O., Hendroff, U., Carolan, C., Rudd, P.M., Treacy, E.P.: IgG N-glycans as potential biomarkers for determining galactose tolerance in Classical Galactosaemia. Mol. Genet. Metab. 105(2), 212–220 (2012)PubMedCrossRefGoogle Scholar
  46. 46.
    Stanta, J.L., Saldova, R., Struwe, W.B., Byrne, J.C., Leweke, F.M., Rothermund, M., Rahmoune, H., Levin, Y., Guest, P.C., Bahn, S., Rudd, P.M.: Identification of N-Glycosylation changes in the CSF and serum in patients with schizophrenia. J. Proteome Res. 9(9), 4476–4489 (2010). doi:10.1021/pr1002356 PubMedCrossRefGoogle Scholar
  47. 47.
    Telford, J.E., Bones, J., McManus, C., Saldova, R., Manning, G., Doherty, M., Leweke, F.M., Rothermundt, M., Guest, P.C., Rahmoune, H.: Anti-psychotic treatment of acute paranoid schizophrenia patients with olanzapine results in altered glycosylation of serum glycoproteins. J Proteome Res. (2012) 11(7):3743–52. doi:10.1021/pr300218h. Accessed 29 May 2012Google Scholar
  48. 48.
    Cosgrave, E.F., Struwe, W.B., Kattla, J.J., Campbell, M.P., Wormald, M.R., Rudd, P.M.: Systems biology glycomics. In: Moo-Young, M. (ed.) Comprehensive Biotehcnology, vol. 1. pp. 427–446. Elsevier (2011).
  49. 49.
    Fernandes, D.: Reducing risk in biopharmaceutical production by controlling Glycosylation. Eur. Biopharm. Rev. 92–97 (2004)Google Scholar
  50. 50.
    Covic, A., Cannata-Andia, J., Cancarini, G., Coppo, R., Frazao, J.M., Goldsmith, D., Ronco, P., Spasovski, G.B., Stenvinkel, P., Utas, C., Wiecek, A., Zoccali, C., London, G.: Biosimilars and biopharmaceuticals: what the nephrologists need to know - a position paper by the ERA-EDTA Council. Nephrol. Dial. Transplant. 23(12), 3731–3737 (2008). doi:10.1093/ndt/gfn519 PubMedCrossRefGoogle Scholar
  51. 51.
    Uçaktürk, E.: Analysis of glycoforms on the glycosylation site and the glycans in monoclonal antibody biopharmaceuticals. J. Sep. Sci. 35(3), 341–350 (2012). doi:10.1002/jssc.201100684 PubMedCrossRefGoogle Scholar
  52. 52.
    Chung, C.H., Mirakhur, B., Chan, E., Le, Q.-T., Berlin, J., Morse, M., Murphy, B.A., Satinover, S.M., Hosen, J., Mauro, D., Slebos, R.J., Zhou, Q., Gold, D., Hatley, T., Hicklin, D.J., Platts-Mills, T.A.E.: Cetuximab-induced anaphylaxis and IgE specific for galactose-α-1,3-Galactose. N. Engl. J. Med. 358(11), 1109–1117 (2008). doi:10.1056/NEJMoa074943 PubMedCrossRefGoogle Scholar
  53. 53.
    Knezevic, A., Polasek, O., Gornik, O., Rudan, I., Campbell, H., Hayward, C., Wright, A., Kolcic, I., O’Donoghue, N., Bones, J., Rudd, P.M., Lauc, G.: Variability, heritability and environmental determinants of human plasma N-glycome. J. Proteome Res. 8(2), 694–701 (2008). doi:10.1021/pr800737u CrossRefGoogle Scholar
  54. 54.
    Lauc, G., Essafi, A., Huffman, J.E., Hayward, C., Knezevic, A., Kattla, J.J., Polasek, O., Gornik, O., Vitart, V., Abrahams, J.L., Pucic, M., Novokmet, M., Redzic, I., Campbell, S., Wild, S.H., Borovecki, F., Wang, W., Kolcic, I., Zgaga, L., Gyllensten, U., Wilson, J.F., Wright, A.F., Hastie, N.D., Campbell, H., Rudd, P.M., Rudan, I.: Genomics meets glycomics the first GWAS study of human N-glycome identifies HNF1A as a master regulator of plasma protein Fucosylation. PLoS Genet. 6(12), e1001256 (2010)PubMedCrossRefGoogle Scholar
  55. 55.
    Fazio, F., Bryan, M.C., Blixt, O., Paulson, J.C., Wong, C.-H.: Synthesis of sugar arrays in microtiter plate. J. Am. Chem. Soc. 124(48), 14397–14402 (2002). doi:10.1021/ja020887u PubMedCrossRefGoogle Scholar
  56. 56.
    Horlacher, T., Seeberger, P.H.: Carbohydrate arrays as tools for research and diagnostics. Chem. Soc. Rev. 37(7), (2008). doi:10.1039/B708016F
  57. 57.
    Disney, M.D., Seeberger, P.H.: The use of carbohydrate microarrays to study carbohydrate-cell interactions and to detect pathogens. Chem. Biol. 11(12), 1701–1707 (2004)PubMedCrossRefGoogle Scholar
  58. 58.
    Laurent, N., Voglmeir, J., Flitsch, S.L.: Glycoarrays-tools for determining protein-carbohydrate interactions and glycoenzyme specificity. Chem. Commun. (37), 4400–4412 (2008). doi:10.1039/B806983M
  59. 59.
    Feizi, T., Chai, W.: Oligosaccharide microarrays to decipher the glyco code. Nat. Rev. Mol. Cell Biol. 5(7), 582–588 (2004)PubMedCrossRefGoogle Scholar
  60. 60.
    Paulson, J.C., Blixt, O., Collins, B.E.: Sweet spots in functional glycomics. Nat. Chem. Biol. 2(5), 238–248 (2006)PubMedCrossRefGoogle Scholar
  61. 61.
    Ratner, D.M., Seeberger, P.H.: Carbohydrate microarrays as tools in HIV glycobiology. Curr. Pharm. Des. 13(2), 173–183 (2007)PubMedCrossRefGoogle Scholar
  62. 62.
    Pedersen, J.W., Blixt, O., Bennett, E.P., Tarp, M.A., Dar, I., Mandel, U., Poulsen, S.S., Pedersen, A.E., Rasmussen, S., Jess, P., Clausen, H., Wandall, H.H.: Seromic profiling of colorectal cancer patients with novel glycopeptide microarray. Int. J. Cancer 128(8), 1860–1871 (2011). doi:10.1002/ijc.25778 PubMedCrossRefGoogle Scholar
  63. 63.
    Lawrie, C.H., Marafioti, T., Hatton, C.S., Dirnhofer, S., Roncador, G., Went, P., Tzankov, A., Pileri, S.A., Pulford, K., Banham, A.H.: Cancer-associated carbohydrate identification in Hodgkin’s lymphoma by carbohydrate array profiling. Int. J. Cancer 118(12), 3161–3166 (2006). doi:10.1002/ijc.21762 PubMedCrossRefGoogle Scholar
  64. 64.
    Park, S., Lee, M.R., Shin, I.: Carbohydrate microarrays as powerful tools in studies of carbohydrate-mediated biological processes. Chem. Commun. (Camb.) (37), 4389–4399 (2008). doi:10.1039/b806699j
  65. 65.
    Hanson, S., Best, M., Bryan, M.C., Wong, C.H.: Chemoenzymatic synthesis of oligosaccharides and glycoproteins. Trends Biochem. Sci. 29(12), 656–663 (2004). doi:10.1016/j.tibs.2004.10.004 PubMedCrossRefGoogle Scholar
  66. 66.
    Seeberger, P.H., Werz, D.B.: Automated synthesis of oligosaccharides as a basis for drug discovery. Nat. Rev. Drug Discov. 4(9), 751–763 (2005). doi:10.1038/nrd1823 PubMedCrossRefGoogle Scholar
  67. 67.
    Feizi, T., Chai, W.: Oligosaccharide microarrays to decipher the glyco code. Nat. Rev. Mol. Cell Biol. 5(7), 582–588 (2004). doi:10.1038/nrm1428 PubMedCrossRefGoogle Scholar
  68. 68.
    Blixt, O., Head, S., Mondala, T., Scanlan, C., Huflejt, M.E., Alvarez, R., Bryan, M.C., Fazio, F., Calarese, D., Stevens, J., Razi, N., Stevens, D.J., Skehel, J.J., van Die, I., Burton, D.R., Wilson, I.A., Cummings, R., Bovin, N., Wong, C.-H., Paulson, J.C.: Printed covalent glycan array for ligand profiling of diverse glycan binding proteins. Proc. Natl. Acad. Sci. U. S. A. 101(49), 17033–17038 (2004). doi:10.1073/pnas.0407902101 PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Tharmala Tharmalingam
    • 1
  • Barbara Adamczyk
    • 1
  • Margaret A. Doherty
    • 1
  • Louise Royle
    • 2
  • Pauline M. Rudd
    • 1
  1. 1.NIBRT Glycobiology Laboratory, NIBRT - The National Institute for Bioprocessing Research and TrainingCo. DublinIreland
  2. 2.Ludger Ltd., Culham Science CentreOxfordshireUK

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