PRODUCTION AND PROPERTIES EVALUATION OF MONOCLONAL ANTIBODIES AGAINST PSEUDOMONAS AERUGINOSA EXOTOXIN A


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Abstract

Aim. Production, study ofproperties and evaluation ofa possibility to use monoclonal antibodies against Pseudomonas aeruginosa exotoxin A (ETA) for the detection ofmolecules ofrecombinant exotoxin A and anatoxin. Materials and methods. Producer hybridomas for monoclonal antibodies against ETA were generated. Recombinant exotoxin A, its atoxic forms, P.aeruginosa anatoxin and P. aeruginosa PA-103 live culture were used for immunization of mice. Results. The experiment of fusion of malignant cell line with immune lymphocytes obtained from a mouse immunized with recombinant ETA turned out to be the most productive. Intensity of interaction of monoclonal antibodies with recombinant atoxic forms ofETA was evaluated in enzyme immunoassay. Protective (toxin-neutralizing) activity of antibodies in cell culture and the ability to detect exotoxin A in latex-agglutination reaction were studied. Antibodies of hybrid culture No. 21 are able to detect molecules of recombinant ETA and anatoxin in the latex-agglutination reaction. Conclusion. Use of the antibodies produced for testing toxigenicity of P. aeruginosa strains as well as detection of anatoxin in the technological process of production of prophylaxis preparations based on exotoxin A is expected.

About the authors

A. V Soldatenkova

Mechnikov Research Institute of Vaccines and Sera, Moscow, Russia

I. V Yakovleva

Mechnikov Research Institute of Vaccines and Sera, Moscow, Russia

N. V Gavrilova

Mechnikov Research Institute of Vaccines and Sera, Moscow, Russia

N. A Mikhailova

Mechnikov Research Institute of Vaccines and Sera, Moscow, Russia

V. V Sviridov

Mechnikov Research Institute of Vaccines and Sera, Moscow, Russia

References

  1. Исаков М.А., Солдатенкова А.В., Калошин А.А., Михайлова Н.А. Иммунобиологические свойства рекомбинантных атоксичных форм экзотоксина А Pseudomonas aeruginosa. Журн. микробиол. 2011, 2: 37-42.
  2. Ломтатидзе М.Н., Храпова Н.П., Крючкова Т.П. и др. Экспресс-выявление Pseudomonas aeruginosa при помощи меченных ФИТЦ моноклональных антител. Эпидемиология и вакциопрофилактика. 2004, 2 (15): 42-45.
  3. Chia J., Pollak M., Avigan D., Steinbch S. Functionally distinct monoclonal antibodies reactive with enzymatically active and binding domains of Pseudomonas aeruginosa toxin A. Infect. Immun. 1986, 52 (3): 756-762.
  4. Danielsen L., Westh H., Balselv E. et al. Pseudomonas aeruginosa exotoxin A antibodies in rapidly deteriorating chronic leg ulcers. Lancet. 1996, 347: 265.
  5. Elzaim H. S., Chopra A. K., Peterson J. W. et al. Protection against exotoxin A (ETA) and Pseudomonas aeruginosa infection in mice with ETA-specific antipeptide antibodies. Infect. Immun. 1998, 66 (11): 5551-5554.
  6. Galloway D. R., Hedstrom R.C., Pavlovskis O.R. Production and characterization of monoclonal antibodies to exotoxin A from Pseudomonas aeruginosa. Infect. Immun. 1984, 44 (2): 262267.
  7. Goding J.W. Monoclonal antibodies: principles and practice. London, Acad. Press. Inc. Ltd, 1986.
  8. Ogata M., Pastan I., Fitzgerald D. Analysis of Pseudomonas exotoxin activation and conformational changes by using monoclonal antibodies as probes. Infect. Immun. 1991, 59 (1): 407-414.
  9. Ohtsuka H., Horigome K., Higuchi A. Binding of monoclonal antibody specific for domain Ia/II of Pseudomonas aeruginosa exotoxin A at pH 4 strongly neutralize exotoxin A-induced cytotoxicity in cell culture and in vivo. Infect. Immun. 1992, 60 (3): 1061-1068.
  10. Shang H., Yfeh M., Lin C., Hwang J. Characterization of monoclonal antibody B7, which neutralizes the cytotoxicity of Pseudomonas aeruginosa exotoxin A. Clin. Diagn. Lab. Immun. 1996, 3 (6): 727-732.
  11. Schmitt C.K., Meysick K.C., O'Brien A.D. Bacterial toxins: friends or foes? Emerg. Infect. Dis. 1999, 5(2): 224-234.

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Copyright (c) 2014 Soldatenkova A.V., Yakovleva I.V., Gavrilova N.V., Mikhailova N.A., Sviridov V.V.

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