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Fabrication and application of carbohydrate microarray for analyzing human serum antibody–carbohydrate interaction

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Abstract

We introduced a strategy for preparing a carbohydrate microarray and demonstrated its utility for characterizing carbohydrate binding and activities. We isolated the lipopolysaccharide (LPS) components from different bacteria and explored the possibility of immobilizing these glycoconjugates on a high-binding polystyrene plate. Carbohydrate-specific combination was examined by observing the binding of the blood group B analogic LPS O-polysaccharide from Escherichia coli on the high-binding polystyrene plate and anti-B from a broad spectra antibody of human blood serum. Strong binding of antibodies was screened, as it was evident that relative response value is two times higher than control. The hybridization results indicated that this method is a reliable technique for the detection of human intestinal bacteria and is expected to be applied in diagnostics and seroepidemiology.

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References

  • Adams EW, Ueberfeld J, Ratner DM, O’Keefe BR, Walt DR, Seeberger PH (2003) Encoded fiber-optic microsphere arrays for probing protein-carbohydrate interactions. Angew Chem Int Ed Engl 42:5317–5320

    Article  CAS  PubMed  Google Scholar 

  • Adams EW, Ratner DM, Bokesch HR, McMahon JB, O’Keefe BR, Seeberger PH (2004) Oligosaccharide and glycoprotein microarrays as tools in HIV glycobiology; glycan-dependent gp120/protein interactions. Chem Biol 11:875–881

    Article  CAS  PubMed  Google Scholar 

  • Ailus K, Palosuo T (1995) IgM class autoantibodies in human cord serum. J Reprod Immunol 29:61–67

    Article  CAS  PubMed  Google Scholar 

  • Andersson M, Carlin N, Leontein K, Lindquist U, Slettengren K (1989) Structural studies of the O-antigenic polysaccharide of Escherichia coli O86, which possesses blood-group B activity. Carbohydr Res 185:211–223

    Article  CAS  PubMed  Google Scholar 

  • Bryan MC, Fazio F, Lee HK, Huang CY, Chang A, Best MD, Calarese DA, Blixt O, Paulson JC, Burton D, Wilson IA, Wong CH (2004) Covalent display of oligosaccharide arrays in microtiter plates. J Am Chem Soc 126:8640–8641

    Article  CAS  PubMed  Google Scholar 

  • Disney MD, Seeberger PH (2004) The use of carbohydrate microarrays to study carbohydrate–cell interactions and to detect pathogens. Chem Biol 11:1701–1707

    Article  CAS  PubMed  Google Scholar 

  • Fukui S, Feizi T, Galustian C, Lawson AM, Chai W (2002) Oligosaccharide microarrays for high-throughput detection and specificity assignments of carbohydrate–protein interactions. Nat Biotechnol 20:1011–1017

    Article  CAS  PubMed  Google Scholar 

  • Galili U, Buehler J, Shohet SB, Macher BA (1987) The human natural anti-Gal IgG. III. The subtlety of immune tolerance in man as demonstrated by crossreactivity between natural anti-Gal and anti-B antibodies. J Exp Med 165:693–704

    Article  CAS  PubMed  Google Scholar 

  • Greenwell P (1997) Blood group antigens: molecules seeking a function? Glycoconj J 14:159–173

    Article  CAS  PubMed  Google Scholar 

  • Houseman BT, Mrksich M (2002) Carbohydrate arrays for the evaluation of protein binding and enzymatic modification. Chem Biol 9:443–454

    Article  CAS  PubMed  Google Scholar 

  • Huang GL, Liu TC, Liu MX, Mei XY (2005) The application of quantum dots as fluorescent label to glycoarray. Anal Biochem 340:52–56

    Article  CAS  PubMed  Google Scholar 

  • Huang GL, Huang HL, Zhang HC, Wang PG (2006a) Structure–function relations of carbohydrates by neoglycolipid arrays. Appl Biochem Biotechnol 133:211–215

    Article  CAS  PubMed  Google Scholar 

  • Huang GL, Zhang HC, Wang PG (2006b) Fabrication and application of neoglycolipid arrays in a microtiter plate. Bioorg Med Chem Lett 16:2031–2033

    Article  CAS  PubMed  Google Scholar 

  • Salles MF, Mandine E, Zalisz R, Guenounou M, Smets P (1989) Protective effects of murine monoclonal antibodies in experimental septicemia: E. coli antibodies protect against different serotypes of E. coli. J Infect Dis 159:641–647

    Article  CAS  PubMed  Google Scholar 

  • Seidl KJ, MacKenzie JD, Wang D, Kantor AB, Kabat EA, Herzenberg LA, Herzenberg LA (1997) Recurrent identical rearrangement and repeated expression of identical heavy and light chains in single anti-phosphatidylcholine B cells. Ann N Y Acad Sci 815:484–488

    Article  CAS  PubMed  Google Scholar 

  • Springer GF (1956) Inhibition of blood-group agglutinins by substances occurring in plants. J Immunol 76:399–407

    Article  CAS  PubMed  Google Scholar 

  • Springer GF, and Horton RE (1969) Blood group isoantibody stimulation in man by feeding blood group-active bacteria. J Clin Invest 48:1280–1291

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Thirumalapura NR, Morton RJ, Ramachandran A, Malayer JR (2005) Lipopolysaccharide microarrays for the detection of antibodies. J Immunol Methods 298:73–81

    Article  CAS  PubMed  Google Scholar 

  • Wang D, Liu S, Trummer BJ, Deng C, Wang A (2002) Carbohydrate microarrays for the recognition of cross-reactive molecular markers of microbes and host cells. Nat Biotechnol 20:275–281

    Article  CAS  PubMed  Google Scholar 

  • Westphal O, Jann K, Himmelspach K (1983) Chemistry and immunochemistry of bacterial lipopolysaccharides as cell wall antigens and endotoxins. Prog Allergy 33:9–39

    CAS  PubMed  Google Scholar 

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Correspondence to Gang-Liang Huang.

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Huang, GL., Mei, XY., Zhang, HX. et al. Fabrication and application of carbohydrate microarray for analyzing human serum antibody–carbohydrate interaction. Appl Microbiol Biotechnol 107 (Suppl 1), 1–7 (2023). https://doi.org/10.1007/s00253-006-0723-9

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  • DOI: https://doi.org/10.1007/s00253-006-0723-9

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