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Broth Microdilution Antibacterial Assay of Peptides

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Peptide Characterization and Application Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 386))

Summary

Native peptides exhibit various biological activities from which the antimicrobial property is one of the most frequently studied. A convenient way of telling whether peptides influence the life cycle of bacteria is the broth microdilution assay. In this measure, growing bacteria are incubated with peptides and growth inhibition is detected with colorimetric methods. Highly charged and protease-sensitive peptides need special considerations in assay design and readout interpretation to reveal the true antimicrobial efficacy and potential utility as human or veterinary therapeutics. The broth microdilution assay is suitable for first assessment of antimicrobial resistance induction.

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References

  1. Loffet, A. (2002) Peptides as drugs: is there a market? J. Pept. Sci. 8, 1–7.

    Article  PubMed  CAS  Google Scholar 

  2. Jones, R. M., and Bulaj, G. (2000) Conotoxins—new vistas for peptide therapeutics. Curr. Pharm. Des. 6, 1249–1285.

    Article  PubMed  CAS  Google Scholar 

  3. Cudic, M. and Otvos, L., Jr. (2002) Intracellular targets of antibacterial peptides. Curr. Drug Targets 3, 101–106.

    Article  PubMed  CAS  Google Scholar 

  4. Zasloff, M. (2002) Antimicrobial peptides of multicellular organisms. Nature 415, 389–395.

    Article  PubMed  CAS  Google Scholar 

  5. Sabath, L. D. (1976) The assay of antimicrobial compounds. Hum. Pathol. 7, 287–185.

    Article  PubMed  CAS  Google Scholar 

  6. Amsterdam, D. (1996) Susceptibility testing of antimicrobials in liquid media, in: Antibiotics in Laboratory Medicine (Loman, V., ed). Williams and Wilkins, Philadelphia, PA: pp. 52–111.

    Google Scholar 

  7. Rahman, M., Kuhn, I., Rahman, M., Olsson-Liljequist, B., and Mollby, R. (2004) Evaluation of a scanner-assisted colorimetric MIC method for susceptibility testing of gram-negative fermentative bacteria. Appl. Environment. Microbiol. 70, 2398–2403.

    Article  CAS  Google Scholar 

  8. Otvos, L., Jr. (2000) Antibacterial peptides isolated from insects. J. Pept. Sci. 6, 497–511.

    Article  PubMed  CAS  Google Scholar 

  9. Stubbings, W. J., Bostock, J. M., Ingham, E., and Chopra, I. (2004) Assessment of a microplate method for determining the post-antibiotic effect in Staphylococcus aureus and Escherichia coli. J. Antimicrob Chemother. 54, 139–143.

    Article  PubMed  CAS  Google Scholar 

  10. Rajagopal, S., Eis, N., and Nickerson, K. W. (2003) Eight gram-negative bacteria are 10,000 times more sensitive to cationic detergents than to anionic detergents. Can. J. Microbiol. 49, 775–779.

    Article  PubMed  CAS  Google Scholar 

  11. Krisher, K. K. and Linscott, A. (1994) Comparison of three commercial MIC systems, E test, fastidious antimicrobial susceptibility panel, and FOX fastidious panel, for confirmation of penicillin and cephalosporin resistance in Streptococcus pneumoniae. J. Clin. Microbiol. 32, 2242–2245.

    PubMed  CAS  Google Scholar 

  12. Virella, G. (1996) Microbiology and Infectious Diseases. Williams and Wilkins, Philadelphia, PA.

    Google Scholar 

  13. National Committee for Clinical Laboratory Standards (1997) Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically, 4th ed. Approved standard M7–A4. National Committee for Clinical Laboratory Standards, Wayne, PA.

    Google Scholar 

  14. Cudic, M., Condie, B. A., Weiner, D. J., et al. (2002) Development of novel antibacterial peptides that kill resistant clinical isolates. Peptides 23, 271–283.

    Article  Google Scholar 

  15. Cudic, M., Lockatell, C. V., Johnson, D. E., and Otvos, L., Jr. (2003) In vitro and in vivo activity of a designed antibacterial peptide analog against uropathogens. Peptides 24, 807–820.

    Article  PubMed  CAS  Google Scholar 

  16. Otvos, L., Jr., Snyder, C., Condie, B., Bulet, P., and Wade, J. D. (2005) Chimeric antimicrobial peptides exhibit multiple modes of action. Int. J. Pept. Res. Ther. 11, 29–42.

    Article  CAS  Google Scholar 

  17. Powell, M. F., Stewart, T., Otvos, L., Jr., et al. (1993) Peptide stability in drug development. II. Effect of single amino acid substitution and glycosylation on peptide reactivity in human serum. Pharm. Res. 10, 1268–1273.

    Article  PubMed  CAS  Google Scholar 

  18. Gennaro, R., Zanetti, M., Benincasa, M., Podda, E., and Miani, M. (2002) Pro-rich antimicrobial peptides from animals: structure, biological functions and mechanism of action. Curr. Pharm. Des. 8, 763–778.

    Article  PubMed  CAS  Google Scholar 

  19. Ge, Y., MacDonald, D. L., Holroyd, K. J., Thornsberry, C., Wexler, H., and Zasloff, M. (1999) In vitro properties of pexiganan, and analog of magainin. Antimicrob. Agents Chemother. 43, 782–788.

    PubMed  CAS  Google Scholar 

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Otvos, L., Cudic, M. (2007). Broth Microdilution Antibacterial Assay of Peptides. In: Fields, G.B. (eds) Peptide Characterization and Application Protocols. Methods in Molecular Biology™, vol 386. Humana Press. https://doi.org/10.1007/978-1-59745-430-8_12

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  • DOI: https://doi.org/10.1007/978-1-59745-430-8_12

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-550-7

  • Online ISBN: 978-1-59745-430-8

  • eBook Packages: Springer Protocols

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