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Characterization of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis

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Abstract

An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. It has been recently suggested that IgG glycosylation defect may be involved in RA immunopathogenesis. Complete characterization of glycans, including both qualitative and quantitative analysis, requires a combination of different techniques, and accurate, robust, sensitive, and high-throughput methodologies are important for analysis of clinical samples. In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. Assignment of the IgG N-glycan structures was made through branching pattern analysis by MSn with high-throughput. Further accurate quantitation indicated that galactosylation and sialylation of IgG N-glycans in RA cases were significantly lower than in healthy subjects. The results indicate that CE coupled with MSn can identify abnormal glycosylation of IgG in RA patients compared with healthy people, and that the present work is useful for RA mechanism studies and RA diagnosis.

Qualitative and quantitative analysis of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis

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References

  1. Axford JS, Sumar N, Alavi A, Isenberg DA, Young A, Bodman KB, Roitt IM. Changes in normal glycosylation mechanisms in autoimmune rheumatic disease. J Clin Invest. 1999;89:1021–31.

  2. Lau KS, Partridge EA, Grigorian A, Silvescu CI, Reinhold VN, Demetriou M, Dennis JW. Complex N-glycan number and degree of branching cooperate to regulate cell proliferation and differentiation. Cell. 2007;129:123–34.

  3. Pang P-C, Chiu PCN, Lee C-L, Chang L-Y, Panico M, Morris HR, Haslam SM, Khoo KH, Clark GF, Yeung WS, Dell A. Human sperm binding is mediated by the Sialyl-Lewis (X) oligosaccharide on the Zona Pellucida. Science. 2011;333:1761–4.

  4. Alley Jr WR, Vasseur JA, Goetz JA, Svoboda M, Mann BF, Matei DE, Menning N, Hussein A, Mechref Y, Novotny MV. N-linked glycan structures and their expressions change in the blood sera of ovarian cancer patients. J Proteome Res. 2012;11:2282–300.

  5. Ohmi Y, Ise W, Harazono A, Takakura D, Fukuyama H, Baba Y, Narazaki M, Shoda H, Takahashi N, Ohkawa Y, Ji S, Sugiyama F, Fujio K, Kumanogoh A, Yamamoto K, Kawasaki N, Kurosaki T, Takahashi Y, Furukawa K. Sialylation converts arthritogenic IgG into inhibitors of collagen-induced arthritis. Nat Commun. 2016;7:11205–16.

  6. Parekh RB, Dwek RA, Sutton BJ, Fernandes DL, Leung A, Stanworth D, Rademacher TW, Mizuochi T, Taniguchi T, Matsuta K. Association of rheumatoid arthritis and primary osteoarthritis with changes in the glycosylation pattern of total serum IgG. Nature (London). 1985;316:452–7.

  7. Field MC, Amatayakui-Chantler S, Rademacher TW, Rudd PM, Dwek RA. Structural analysis of the N-glycans from human immunoglobulin A1: comparison of normal human serum immunoglobulin A1 with that isolated from patients with rheumatoid arthritis. Biochem J. 1994;299:261–75.

    Article  CAS  Google Scholar 

  8. Rademacher TW, Williams P, Dwek RA. Agalactosyl glycoforms of IgG autoantibodies are pathogenic. Proc Natl Acad Sci. 1994;91:6123–7.

    Article  CAS  Google Scholar 

  9. Malhotra R, Wormald MR, Rudd PM, Fischer PB, Dwek RA, Sim RB. Glycosylation changes of IgG associated with rheumatoid arthritis can activate complement via the mannose-binding protein. Nat Med. 1995;1:237–43.

    Article  CAS  Google Scholar 

  10. Youings A, Chang SC, Dwek RA, Scragg IG. Site-specific glycosylation of human immunoglobulin G is altered in four rheumatoid arthritis patients. Biochem J. 1996;314:621–30.

    Article  Google Scholar 

  11. Dwek RA, Lellouch AC, Wormald MR. Glycobiology: the function of the sugar in the IgG molecule. J Anat. 1995;187:279–92.

    CAS  Google Scholar 

  12. Hänsler M, Kötz K, Häntzschel H. Detection of immunoglobulin G glycosylation changes in patients with rheumatoid arthritis by means of isoelectric focusing and lectin-affinoblotting. Electrophoresis. 1995;16:811-2.

  13. Gindzienska-Sieskiewicz E, Radziejewska I, Domysławska I, Klimiuk PA, Sulik A, Rojewska J, Gabryel-Porowska H, Sierakowski S. Changes of glycosylation of IgG in rheumatoid arthritis patients treated with methotrexate. Adv Med Sci. 2016;61:193–7.

  14. Pekelharing JM, Hepp E, Kamerling JP, Gerwig GJ, Leijnse B. Alterations in carbohydrate composition of serum IgG from patients with rheumatoid arthritis and from pregnant women. Ann Rheum Dis. 1988;47:91–5.

    Article  CAS  Google Scholar 

  15. North SJ, Huang HH, Sundaram S, Jang-Lee J, Etienne AT, Trollope A, Chalabi S, Dell A, Stanley P, Haslam SM. Glycomics profiling of Chinese hamster ovary cell glycosylation mutants reveals N-glycans of a novel size and complexity. J Biol Chem. 2010;285:5759–75.

  16. Yu SY, Wu SW, Khoo KH. Distinctive characteristics of MALDI-Q/TOF and TOF/TOF tandem mass spectrometry for sequencing of permethylated complex type N-glycans. Glycoconj J. 2006;23:355–69.

    Article  Google Scholar 

  17. Huang C, Zhan T, Liu Y, Li Q, Wu H, Ji D, Li Y. Glycomic profiling of carcinoembryonic antigen isolated from human tumor tissue. Clin Proteomics. 2015;12:17.

  18. Yu G, Zhao X, Yang B, Ren S, Guan H, Zhang Y, Lawson AM, Chai W. Sequence determination of sulfated carrageenan-derived oligosaccharides by high-sensitivity negative-ion electrospray tandem mass spectrometry. Anal Chem. 2006;78:8499–505.

  19. Bondt A, Rombouts Y, Selman MHJ, Hensbergen PJ, Reiding KR, Hazes JM, Dolhain RJ, Wuhrer M. Immunoglobulin G (IgG) Fab glycosylation analysis using a new mass spectrometric high-throughput profiling method reveals pregnancy-associated changes. Mol Cell Probes. 2014;13:3029–39.

  20. Ashline DJ, Yu Y, Lasanajak Y, Song X, Hu L, Ramani S, Prasad V, Estes MK, Cummings RD, Smith DF, Reinhold VN. Structural characterization by multistage mass spectrometry (MSn) of human milk glycans recognized by human rotaviruses. Mol Cell Probes. 2014;13:2961–74.

  21. Yang S, Li Y, Shah P, Zhang H. Glycomic analysis using glycoprotein immobilization for glycan extraction. Anal Chem. 2013;85:5555–61.

    Article  CAS  Google Scholar 

  22. Rillahan CD, Antonopoulos A, Lefort CT, Sonon R, Azadi P, Ley K, Dell A, Haslam SM, Paulson JC. Global metabolic inhibitors of sialyl- and fucosyltransferases. Nat Chem Biol. 2016;8:661–8.

  23. Bond A, Alavi A, Axford JS, Bourke BE, Brukner FE, Kerr MA, Maxwell JD, Tweed KJ, Weldon MJ, Youinou P, Hay FC. A detailed lectin analysis of IgG glycosylation, demonstrating disease specific changes in terminal galactose and N-acetylglucosamine. J Autoimmun. 1997;10:77–85.

  24. Pasek M, Duk M, Podbielska M, Sokolik R, Szechinski J, Lisowska E, Krotkiewski H. Galactosylation of IgG from rheumatoid arthritis (RA) patients: changes during therapy. Glycoconj J. 2006;23:463–71.

  25. Weiz S, Wieczorek M, Schwedler C, Kaup M, Braicu EI, Sehouli J, Tauber R, Blanchard V. Acute-phase glycoprotein N-glycome of ovarian cancer patients analyzed by CE-LIF. Electrophoresis. 2016;37:1461–7.

  26. Zaia J. Capillary electrophoresis-mass spectrometry of carbohydrates. Methods Mol Biol. 2013;984:13–25.

    Article  CAS  Google Scholar 

  27. Canis K, Mxkinnon TA, Nowak A, Haslam SM, Panico M, Morris HR, Laffan MA, Dell A. Mapping the N-glycome of human Von Willebrand factor. Biochem J. 2012;447:217–28.

  28. Wang C-Y, Hsieh Y-Z. Analysis of chitin oligosaccharides by capillary electrophoresis with laser-induced fluorescence. J Chromatogr A. 2002;979:431–8.

    Article  CAS  Google Scholar 

  29. Ruhaak LR, Hennig R, Huhn C, Borowiak M, Dolhain RJ, Deelder AM, 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. 2010;9:6655–64.

  30. Pitt JJ, Gorman JJ. Oligosaccharide characterization and quantitation using 1-Phenyl-3-methyl-5-pyrazolone derivatization and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Anal Biochem. 1997;248:63–75.

    Article  CAS  Google Scholar 

  31. Tarelli E, Byers HL, Wilson M, Roberts G, Homer KA, Beighton D. Detecting mannosidase activities using ribonuclease B and matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Anal Biochem. 2000;282:165–72.

    Article  CAS  Google Scholar 

  32. Gonzalez L, Bruix M, Diaz-Maurino T, Ten F, Rico M, Solis D, Jiménez-Barbero J. Conformational studies of the Man8 oligosaccharide on native ribonuclease B and on the reduced and denatured protein. J Arch Biochem Biophys. 2000;383:17–27.

  33. Yamashita K, Totani K, Kuroki M, Matsuoka Y, Ueda I, Kobata A. Structural studies of the carbohydrate moieties of carcinoembryonic antigens. Cancer Res. 1987;47:3451–9.

    CAS  Google Scholar 

  34. Chandrasekaran EV, Davila M, Nixon DW, Goldfarb M, Mendicino J. Isolation and structures of the oligosaccharide units of carcinoembryonic antigen. J Biol Chem. 1983;258:7213–22.

    CAS  Google Scholar 

  35. Detl Z, Miksfk I, Charvatova J, Eckhardt A. Comparison of the electrophoretic separation of proteins in capillaries with different inner diameter. J Chromatogr A. 2003;1013:233–8.

  36. Fu D, Chen L, O’Neil RA. A detailed structural characterization of ribonuclease B oligosaccharides by 1H NMR spectroscopy and mass spectrometry. Carbohydr Res. 1994;261:173–86.

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Acknowledgments

This work was supported by the Scientific Equipment Development of Chinese Academy of Sciences (No.YZ201249) and National High-tech Research and Development Projects (863 Program, No. 2014AA021101). We are also indebted to the Natural Science Foundation of China (No. 31270909) and Major State Basic Research Development Program of China (973 Program, No. 2013CB910100), Great help from Torsten Juelich in Peking University for assistance with discussion and editing is greatly acknowledged.

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Authors and Affiliations

  1. Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing, 100101, China

    Chuncui Huang, Yaming Liu, Hongmei Wu, Dehui Sun & Yan Li

  2. GuangDong Bio-Healtech Advanced Co., Ltd, Foshan, Guangdong, 528000, China

    Hongmei Wu

  3. University of Chinese Academy of Sciences, 19 A Yuquan Rd, Shijingshan District, Beijing, 100049, China

    Yan Li

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  1. Chuncui Huang
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Correspondence to Yan Li.

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Human and Animal Rights and Informed Consent

All procedures performed in studies involving human participants were in accordance with the ethical standards of Biosafety and Ethics Committee in Institute of Biophysics, Chinese Academy of Sciences, and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

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Informed consent was obtained from all the patients and healthy subjects included in the study.

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Huang, C., Liu, Y., Wu, H. et al. Characterization of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. Anal Bioanal Chem 409, 3731–3739 (2017). https://doi.org/10.1007/s00216-017-0302-1

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  • DOI: https://doi.org/10.1007/s00216-017-0302-1

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