Effects of epitope sequence tandem repeat and proline incorporation on polyclonal antibody production against cytochrome 1A2 and 3A4

  • Ahn, Tae-Ho (Department of Biochemistry, College of Veterinary Medicine, Chonnam National University) ;
  • Yun, Chul-Ho (School of Biological Sciences and Technology, Chonnam National University)
  • Published : 2009.07.31


We describe a method for producing polyclonal antibodies against peptide antigen cytochrome P450 1A2 and 3A4 using a tandem repeat of the epitope region and incorporation of proline residue between the repeated sequences. An ELISA assay revealed more efficient generation of polyclonal antibodies to tandem repeat peptide antigens than mono-epitope peptides. The incorporation of proline residues further stimulated antibody production.


  1. Angeletti, R. H. (1999) Design of useful peptide antigens. J. Biomol. Tech. 10, 2-10
  2. Edwards, R. J., Singleton, A. M., Murray, B. P., Davies, D. S. and Boobis, A. R. (1995) Short synthetic peptides exploited for reliable and specific targeting of antibodies to the C-termini of cytochrome P450 enzymes. Biochem. Pharmacol. 49, 39-47
  3. Wang, R. W., Newton, D. J., Liu, N. Y., Shou, M., Rushmore, T. and Lu, A. Y. (1999) Inhibitory anti-CYP3A4 peptide antibody: mapping of inhibitory epitope and specificity toward other CYP3A isoforms. Drug. Metab. Dispos. 27, 167-172
  4. Kim, M., Yun, C-H., Park, S. K., Seo, J. H. and Ahn, T. (2007) Production of polyclonal antibodies against peptide antigens using polystyrene beads as a carrier. Biotechol. Lett. 29, 1735-1740
  5. Combet, C., Blanchet, C., Geourjon, C. and Deléage, G. (2000) NPS@: network protein sequence analysis. Trends Biochem. Sci. 25, 147-150
  6. Gurunath, R., Beena, T. K., Adiga, P. R. and Balaram, P. (1995) Enhancing peptide antigenicity by helix stabilization. FEBS Lett. 361, 176-178
  7. Subramanian, S., Karande, A. A. and Adiga, P. R. (2001) Helix stabilization in the C-terminal peptide of chicken riboflavin carrier protein enhances immunogenicity and prolongs contraceptive potential as an epitope-based vaccine in female rats. Biochem. Biophys. Res. Commun. 14, 236-243
  8. Reichlin, M. (1980) Use of glutaraldehyde as a coupling agent for proteins and peptides. Methods Enzymol. 70, 159-165
  9. Castell, J. V., Cervera, M. and Marco, R. (1979) A convenient micromethod for the assay of primary amines and proteins with fluorescamine. A re-examination of the conditions of reaction. Anal. Biochem. 99, 379-391