Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지

Use of Antibody Displayed Phage for the Detection of Dextran Using a Dipstick Assay and Transmission Electron Micrograph

  • Kim Du-Woon (Department of Food and Nutrition, Chonnam National University) ;
  • Day Donal F. (The Audubon Sugar Institute, LSU Agricultural Experiment Station)
  • Published : 2006.08.01
Download PDF
⟨ Previous Next ⟩

Abstract

An antibody displayed phage collection (SBAE-2R), screened from a human synthetic phage antibody library (Fab 21ox), was used for the determination of dextran. The dextran-binding affinity was determined by serologically specific transmission electron microscopy (TEM) and a paper dipstick assay. The phage collection was distributed over the dextrancoated grids with 39$\pm$25 phages/$\mu$m$^2$ on the grids. Phages were not seen on dextran-coated grids exposed to the Fab 2lox phage library. The phage collection (SBAE-2R) produced 54$\pm$3 color normalized intensity (N.I.) from 125 ppm to 1,000 ppm of dextran and 5$\pm$1 (N.I.) for 63 ppm of dextran in a paper dipstick assay. This research extends the analytical options for dextran analysis by antibody displayed phage with a minimum of equipment usage.

Keywords

References

  1. Bjorling, E., E. V. Garrelts, A. Morner, M. Ehnlund, and M. A. A. Persson. 1999. Human neutralizing human immunodeficiency virus type-2 specific Fab molecules generated by phage display. J. Gen. Virol. 80: 1987-1993 https://doi.org/10.1099/0022-1317-80-8-1987
  2. Breitling, F. and S. Dubel. 1999. Recombinant Antibody, pp. 2-37. John Wiley & Sons, Inc., New York
  3. Chames, P. and D. Baty. 2000. Antibody engineering and its applications in tumor targeting and intracellular immunization. FEMS Microbiol. Lett. 189: 1-8 https://doi.org/10.1111/j.1574-6968.2000.tb09197.x
  4. Choi, J. H., J. I. Choi, and S. Y. Lee. 2005. Display of proteins on the surface of Escherichia coli by C-terminal deletion fusion to the Salmonella typhimurium OmpC. J. Microbial. Biotechnol. 15: 141-146
  5. Clackson, T., H. R. Hoogenboom, A. D. Griffiths, and G. Winter. 1991. Making antibody fragments using phage display libraries. Nature 352: 624-628 https://doi.org/10.1038/352624a0
  6. Clarke, M. A. and M. A. Godshall. 1988. Determination of dextrans in raw cane sugar by Roberts copper method: Collaborative study. J Assoc. Off. Anal. Chem. 71: 276-279
  7. Deng, S. J., C. R. Mackenzie, J. Sadowska, J. Michniewicz, N. M. Young, D. R. Bundle, and S. A. Narang. 1994. Selection of antibody single-chain variable fragments with improved carbohydrate binding by phage display. J. Biol. Chem. 269: 9533-9538
  8. Derrick, K. S. and R. H. Brlansky. 1976. Assay for viruses and mycoplasmas using serologically specific electron microscopy. Phytopathology 66: 815-820 https://doi.org/10.1094/Phyto-66-815
  9. Dewey, F. M., M. M. MacDonald, and S. I. Phillips. 1989. Development of monoclonal-antibody-ELISA, dot-blot and dip-stick immunoassays for Hurnicola langinosa in rice. J. Gen. Microbiol.135: 361-374
  10. Harrison, J. L., S. C. Williams, G. Winter, and A. Nissim. 1996. Screening of phage antibody libraries. Methods Enzyrnol. 267: 83-109 https://doi.org/10.1016/S0076-6879(96)67007-4
  11. Hwang, O. H., H. G. Park, E. H. Park, S. H. Park, and W. M. Park. 2004. Production of recombinant proteins as immunoanalytical markers of genetically-modified organisms (GMO). J. Microbiol. Biotechnol. 14: 783-788
  12. Kim, D. W. and D. F. Day. 2005. A dip-stick assay for dextran analysis. Proc. Int. Soc. Sugar Cane Technol. 25: 89-92
  13. Kim, J. N., M. J. Seo, E. A. Cho, S. J. Lee, S. B. Kim, C. I. Cheigh, and Y. R. Pyun. 2005. Screening and characterization of an esterase from a metagenomic library. J. Microbiol. Biotechnol. 15: 1067-1072
  14. Medical Research Council. 1995. Protocol for Use of the Human Synthetic Phage-Antibody Fab 2lox Library (Lox Library). Cambridge Centre for Protein Engineering, Cambridge, England
  15. Sanden, G. N., P. K. Cassiday, and J. M. Barbaree. 1993. Rapid immunodot technique for identifying Bordetella pertussis. J. Clin. Microbiol. 31: 170-172
  16. Shim, Y. Y., D. H. Shon, B. Y. Kwak, J. Yu, and K. M. Chee. 2004. Comparison of properties of polyclonal anti-Nacetylchitooligosaccharides antibodies produced for ELISA. J. Microbiol. Biotechnol. 14: 686-692
  17. Towbin, H. and J. Gordon. 1984. Immunoblotting and dot immunobinding - current status and outlook. J. Immunol. Methods 72: 313-340 https://doi.org/10.1016/0022-1759(84)90001-2