Comparison of Properties of Polyclonal Anti-N-Acetylchitooligosaccharides and Anti-Chitooligosaccharides Antibodies Produced for ELISA

  • Shim, Youn-Young (Korea Food Research Institute/Graduate School of Biotechnology, Korea University) ;
  • Shon, Dong-Hwa (Korea Food Research Institute) ;
  • Kwak, Bo-Yeon (Korea Food Research Institute) ;
  • Yu, Jae-Hoon (Life Science Division, Korea Institute of Science and Technology) ;
  • Chee, Kew-Mahn (Graduate School of Biotechnology, Korea University)
  • Published : 2004.08.01

Abstract

To develop the enzyme-linked immunosorbent assay (ELISA) for the analysis of N-acetylehitooligosaccharides (NACOS) and chitooligosaccharides (COS), specific antibodies (Abs) were produced, and their properties were compared. N-acetylehitohexaose (NACOS6), chitohexaose (COS6), and COS mixture (COSM) conjugated to bovine serum albumin (BSA) were used to immunize rabbits. By the use of specific Abs and NACOS6-horseradish peroxidase (HRP), COS6-HRP, and COSM-HRP conjugates, competitive direct ELISA (cdELISA) was developed. The detection limits of NACOS6 by the anti-NACOS6 Ab and COS6 by the anti-COS6 and the anti-COSM Abs in the cdELISAs were about 0.2, 2, ana 2 ng/ml (ppb), respectively. In the cdELISA, the anti-NACOS6 Ab was found to recognize NACOS3-NACOS6, but not N-acetyl-D-glucosamine (GlcNAc), NACOS2, and COSs; the anti-COS6 Ab recognized COS2-COS6 and COSM, but not glucosamine (GlcN) and NACOSs. The recognition pattern of the anti-COSM Ab was almost the same as that of the anti- COS6 Ab, except that the former recognized COS2 and COS3 slightly better than the latter.

Keywords

References

  1. Protein Method Gel electrophoresis under denaturing conditions Bollag, D. M.;M. D. Rozycki;S. J. Edelstein
  2. Nature v.243 Synthetic of a matrix-supported enzyme in non-aqueous conditions Bartling, G. J.;H. D. Brown;S. K. Chattopadhyay
  3. Meth. Enzymol. v.135 Immunobilization techniques for enzymes Hearn, M. T. W. https://doi.org/10.1016/0076-6879(87)35068-2
  4. Advances in Chitin Science Applications of chitin and chitosan as fiber and textile chemicals Hudson, S. M.;A. Domard(ed.);G. A. F.Roberts(ed.);K. M. Varum(ed.)
  5. J. Microbiol. Biotechnol. v.12 Antituor activity of chitosan oligosaccharides produced in ultrafiltration membrane reactor system Jeon, Y. J.;S. K. Kim
  6. Exp. Mycology v.8 Characterization of the smallest chitosan oligomer that is maximally antifungal to Fusrium solani and elicits pisatin formation in Pisum sativum Kendra, D.;L. Hadwiger https://doi.org/10.1016/0147-5975(84)90013-6
  7. Physiol. Mol. Plant Pathol. v.35 Chitosan oligomers from Fusarium solani/pea interactions, chitinase/-glucanase digestion of sporelings and from fungal wall chitin actively inhibit fungal growth and enhance disease resistance Kendra, D. F.;D. Christian;L. A. Hadwiger https://doi.org/10.1016/0885-5765(89)90052-0
  8. Biosci. Biotechnol. Biochem. v.64 Enzyme-linked immunosorbent assay for detection of chitooligosaccharides Kim, S. Y.;D. H. Shon;K. H. Lee https://doi.org/10.1271/bbb.64.696
  9. Food Technology v.1 Recovery and utilization of chitin and chitosan in food processing waste management Knorr, D.
  10. J. Microbiol. Biotechnol. v.13 Detection of Fusarium species by enzyme-linked immunosorbent assay using monoclonal antibody Kwak, B. Y.;B. J. Kwon;C. H. Kweon;D. H. Shon
  11. J. Microbiol. Biotechnol. v.14 Detection of Aspergillus, Penicillium, and fusarium species by sandwich enzyme-linked immunosorbent assay using mixed monoclonal antibodies Kwak, B. Y.;B. J. Kwon;C. H. Kweon;D. H. Shon
  12. Arch. Biochem. Biophys. v.175 Aldonate coupling, a simple procedure for the preparation of carbohydrate-protein conjugates for studies of carbohydratebinding proteins Lonngren, J.;I. J. Goldstein;J. E. Neiderhuber https://doi.org/10.1016/0003-9861(76)90557-9
  13. J. Biol. Resp. Modif v.3 Effect of D-glucosamine on human natural killer activity in vitro Matheson, D. S.;B. J. Green;S. J. Friedman
  14. Vaccine v.4 Stimulation of cytokine production in mice using deacetylatd chitin Nishimura, K.;C. Ishihara;S. Ukei;S. Tokura;I. Azuma https://doi.org/10.1016/0264-410X(86)90002-2
  15. J. Microbiol. Biotechnol. v.12 Variation of antifungal activities of chitosans on plant pathogens Park, R. D.;K. J. Jo;Y. Y. Jo;Y. L. Jin;K. Y. Kim;J. H. Shim;Y. W. Kim
  16. Biochem. J. v.61 The determination of glucosamine and galactosamine Rondle, C. J. M.;W. T. J. Morgan https://doi.org/10.1042/bj0610586
  17. Chem. Pharm. Bull. v.33 Enhancing effect of N-acetylchitooligosaccharides on the active oxygen-generating and microbicidal activitites of peritoneal exudate cells in mice Suzuki, K.;A. Tokoro;Y. Okawa;S. Suzuki;M. Suzuki https://doi.org/10.1248/cpb.33.886
  18. Carbohydr. Res. v.151 Antitumor effect of hexa-N-acetylchitohexaose and chitohexaose Suzuki, K.;T. Mikami;Y. Okawa;A. Tokoro;S. Suzuki;M. Suzuki https://doi.org/10.1016/S0008-6215(00)90359-8
  19. Chem. Pharm. Bull. v.36 Growth-inhibitory effect of hexa-N-acetylchitohexaose and chitohexaose against Meht-A solid tumor Tokoro, A.;N. Tatewaki;K. Suzuki;T. Mikami;S. Suzuki;M. Suzuki https://doi.org/10.1248/cpb.36.784
  20. Bull. Fac. Agr. Sage Univ. v.66 Purification and enzymatic properties of chitosanase from Bacillus sp. No.7-M Uchida, Y.;M. Izume;A. Ohtakara
  21. J. Gen. Appl. Microbiol. v.34 Purification and properites of chitosanase from Bacillus circulans MH-K1 Yabuki, M.;A. Uchiyama;K. Suzuki;A. Ando;T. Fujii https://doi.org/10.2323/jgam.34.255