Production and characterization of anti-Salmonella polyclonal antibodies as bio-recognition element for developing a microbial monitoring method

미생물학적 모니터링 분석방법 개발을 위한 생물학적 수용체로서 살모넬라에 특이적인 다중클론 항체의 생산 및 특성 검토

  • Park, Mi-Kyung (School of Food Science and Biotechnology / Food and Bio-industry Research Institute, Kyungpook National University)
  • 박미경 (경북대학교 식품공학부/식품생물산업 연구소)
  • Received : 2017.11.14
  • Accepted : 2017.11.25
  • Published : 2017.11.30


For the construction of the microbial monitoring method, anti-Salmonella polyclonal antibodies (pAbs) were produced from a rabbit and purified by saturated ammonium sulfate precipitation and protein A affinity column. The reactivity of anti-Salmonella pAbs was compared to that of commercial ones by using an indirect ELISA. The specificity of anti-Salmonella pAbs was investigated using 20 Salmonella serotypes and 20 non-Salmonella strains. A capturing ability of anti-Salmonella pAbs was investigated by exposing antibody-immobilized gold biosensor to different concentration of Salmonella mixture. Anti-Salmonella pAbs were successfully produced and purified with an antibody concentration of 2.0 mg/mL The reactivity of purified anti-Salmonella pAbs was greater than that of commercial one at all tested concentrations. All Salmonella serotypes, except S. Diarizonae, showed excellent binding efficiency with purified anti-Salmonella pAbs. Moreover, the purified anti-Salmonella pAbs showed excellent specificity against all non-Salmonella strains. The anti-Salmonella pAbs immobilized on the gold biosensor demonstrated the successful capturing capability against Salmonella with a dose-response manner. Therefore, the anti-Salmonella pAbs exhibited sufficient reactivity, specificity, as well as capturing capability against Salmonella to be considered as a bio-recognition element.


  1. Mead PS, Slutsker L, Dietz V, McCaig LF, Bresee JS, Shapiro C, Griffin PM, Tauxe RV (1999) Food-related illness and death in the United States. Emerging Infect Dis, 5, 607-625
  2. Tirado C, Schmidt K (2001) WHO Surveillance program for control of foodborne infections and intoxications: preliminary results and trends across greater Europe. J Infect, 43, 80-84
  3. Centers for Disease Control and Prevention (2006) Surveillance for foodborne-disease outbreaks in the United States, 1998-2002. Surveilance summeries, MMWR, 55, 1-34
  4. Nataro JP, Bopp CA, Fields PI, Kaper JB, Strockbine NA (2011) Escherichia, Shigella, and Salmonella. Versalovic J, Carroll KC, Funke G, Jorgensen JH, Landry ML, Warnock DW, ASM press., Washington DC, USA, p 354-383
  5. Currie A, MacDougall L, Aramini J, Gaulin C, Ahmed R, Isaacs S (2005) Frozen chicken nuggets and strips and eggs are leading risk factors for Salmonella Heidelberg infections in Canada. Epidemiol Infect, 133, 809-816
  6. Park MK, Oh JH, Chin BA (2011) The effect of incubation temperature on the binding of Salmonella typhimurium to phage-based magnetoelastic biosensors. Sens Actuators B Chem, 160, 1427-1433
  7. Centers for Disease Control and Prevention. (accessed 2013)
  8. Centers for Disease Control and Prevention. (accessed 2004)
  9. Ricci F, Volpe G, Micheli L, Palleschi G (2007) A review on novel developments and applications of immunosensors in food analysis. Analytica Chimica Acta, 605, 111-129
  10. Su L, Jia W, Hou C, Lei Y (2011) Microbial biosensors: a review. Biosens Bioelectron, 26, 1788-1799
  11. Turner PF (2013) Biosensors: sense and sensibility. Chem Soc Rev, 42, 3184-3196
  12. Sin ML, Mach KE, Wong PK, Liao JC (2014) Advances and challenges in biosensor-based diagnosis of infectious diseases. Expert Rev Mol Diagn, 14, 225-244
  13. Park MK, Park JW, Oh JH (2012) Optimization and application of a dithiobis-succinimidyl propionate-modified immunosensor platform to detect Listeria monocytogenes in chicken skin. Sens Actuators B Chem, 171-172, 323-331
  14. Park MK, Oh JH (2014) Immunomagnetic bead separation coupled with a dithiobis-succinimidyl propionate (DSP)-modified immunosensor to detect Listeria monocytogenes in chicken skin. J Electrochem Soc, 161, B237-B242
  15. Park MK, Oh JH (2012) Rapid detection of Escherichia coli O157:H7 on turnip greens using a modified gold biosensor combined with light microscopic imaging system. J Food Sci, 77, M127-M134
  16. Zeng X, Shen Z, Mernaugh R (2012) Recombinant antibodies and their use in biosensors. Anal Bioanal Chem, 402, 3027-3038
  17. Borrebaeck CA (2000) Antibodies in diagnostics-from immunoassays to protein chips. Immunol Today, 21, 379-381
  18. Deshpande SS (1996) Enzyme immunoassays from concept to product development. Springer International Publishing AG, Boston, USA, p 24-154
  19. Plaza G, Ulfig K, Tien AJ (2000) Immunoassays and environmental studies. Pol J Environ Stud, 9, 231-236
  20. Skottrup PD, Nicolaisen M, Justesen AF (2008) Towards on-site pathogen detection using antibody-based sensors. Biosens Bioelectron, 24, 339-348
  21. Danczyk R, Krieder B, North A, Webster T, HoganEsch H, Rundell A (2003) Comparison of antibody functionality using different immobilization methods. Biotechnol Bioeng, 84, 215-223