medigraphic.com
ISSN 1027-2852 (Electronic)
ISSN 0864-4551 (Print)

2016, Number 1

<< Back Next >>

Biotecnol Apl 2016; 33 (1)

Validation of a sandwich ELISA for the quantification of the main toxin (Pt) of Bordetella pertussis

Lemos-Pérez G, Delgado-Espina M, Quintana-Vazquez D, Alvarez-Acosta A, Coizeau-Rodríguez E, Caballero-Menéndez E

Language: English
References: 36
Page: 1301-1306
PDF size: 495.90 Kb.


ABSTRACT

Bordetella pertussis bacterium causes a highly contagious respiratory disease named Pertussis or whooping cough, which represents a major threat to public health. Pt is the main toxin of B. pertussis and one of the most important virulence factors. The inactivated toxin is considered the major protective antigen in all the new acellular vaccines available today. In this work, a previously established sandwich capture ELISA test for a genetically detoxified Pertussis toxin (Ptg) was validated, with the aim of monitoring the expression of Ptg in bacterial cell culture, and to quantify the antigen throughout the purification process. The validation of this method is a requirement of the pharmaceutical industry; therefore, extensive experiments were performed in order to employ the assay as an analytical tool. The intra- and inter-assay precision was tested and proved a robust starting platform for the assay. The repeatability for the assay was determined using a calibration curve and spiked samples. The assay had a coefficient of variation (CV) of less than 10 %. The inter-assay precision (CV) was lower than 20 % for the calibration curve and lower than 10 % for control samples. Dilution linearity and parallelism were demonstrated. Accuracy (spike recovery) for all concentrations was shown in the range 80-120 %. This assay is highly accurate and reproducible in determining the levels of Ptg in spiked samples, fulfilling the most stringent acceptance criteria and being adequate for the intended analytical purpose. It is currently being applied as an analytical tool in production process development of pertussis antigens for vaccine candidates.


REFERENCES

  1. Bordet J, Gengou D. Le microbe de la coqueluche. Ann Inst Pasteur. 1906;20:731.

  2. Black RE, Cousens S, Johnson HL, Lawn JE, Rudan I, Bassani DG, et al. Global, regional, and national causes of child mortality in 2008: a systematic analysis. Lancet. 2010;375:1969.

  3. Mata AF, Sarnaik AA. Bronchoscopy with N-acetylcysteine lavage in severe respiratory failure from pertussis infection. Pediatrics. 2013;132(5):e1418-23.

  4. Greco D, Salmaso S, Mastrantonio P, Giuliano M, Tozzi AE, Anemona A, et al. A controlled trial of two acellular vaccines and one whole-cell vaccine against pertussis. Progetto Pertosse Working Group. N Engl J Med. 1996;334(6):341-8.

  5. Gustafsson L, Hallander HO, Olin P, Reizenstein E, Storsaeter J. A controlled trial of a two-component acellular, a five-component acellular, and a wholecell pertussis vaccine. N Engl J Med. 1996;334(6):349-55.

  6. Simondon F, Preziosi MP, Yam A, Kane CT, Chabirand L, Iteman I, et al. A randomized double-blind trial comparing a two-component acellular to a whole-cell pertussis vaccine in Senegal. Vaccine. 1997;15(15):1606-12.

  7. Preston A, Maskell DJ. A new era of research into Bordetella pertussis pathogenesis. J Infect. 2002;44(1):13-6.

  8. Hewlett EL, Cherry JD. New and improved vaccines against pertussis. In: Myron ML, Graeme CW, James BK, Gary SC (eds). New generation vaccines. 2nd Ed. New York: Marcel Dekker; 1997. p. 387-416.

  9. Juretzko P, von Kries R, Hermann M, Wirsing von Konig CH, Weil J, Giani G. Effectiveness of acellular pertussis vaccine assessed by hospital-based active surveillance in Germany. Clin Infect Dis. 2002;35(2):162-7.

  10. Zhang L, Prietsch SO, Axelsson I, Halperin SA. Acellular vaccines for preventing whooping cough in children. Cochrane Database Syst Rev. 2011;(1):CD001478.

  11. Farizo KM, Burns DL, Finn TM, Gruber MF, Pratt RD. Clinical evaluation of pertussis vaccines: US Food and Drug Administration regulatory considerations. J Infect Dis. 2014;209 Suppl 1:S28-31.

  12. Moraga-Llop FA, Campins-Martí M. Vacuna de la tos ferina. Reemergencia de la enfermedad y nuevas estrategias de vacunación. Enferm Infecc Microbiol Clin. 2015;33(3):190-6.

  13. Ozcengiz E, Kilinc K, Buyuktanir O, Gunalp A. Rapid purification of pertussis toxin (PT) and filamentous hemagglutinin (FHA) by cation-exchange chromatography. Vaccine. 2004;22(11-12):1570-5.

  14. Tamura M, Nogimori K, Yajima M, Ase K, Ui M. A role of the B-oligomer moiety of islet-activating protein, pertussis toxin, in development of the biological effects on intact cells. J Biol Chem. 1983;258(11):6756-61.

  15. Locht C. Molecular aspects of Bordetella pertussis pathogenesis. Int Microbiol. 1999;2(3):137-44.

  16. Robbins JB, Schneerson R, Kubler-Kielb J, Keith JM, Trollfors B, Vinogradov E, et al. Toward a new vaccine for pertussis. Proc Natl Acad Sci USA. 2014;111(9):3213-6.

  17. Lemos G, Delgado M, Quintana- Vázquez D, Carpio-Muñoz E, Caballero E, Alvarez Acosta A. Development of sandwich ELISA-based assays for screening Bordetella pertussis antigens expressed in the bacterial cell culture. Biotecnol Apl. 2015;32(3):3311-7.

  18. Quintana-Vázquez D, Delgado M, Lemos G, Coizeau E, Ramos Y, Támbara Y, et al. Assessment of the Bordetella pertussis BpCNIC0311 strain as a producing strain of genetically detoxified Toxoid (PTg), filamentous hemagglutinin (FHA) and type 2 Pertactin (Prn2). J Infect Dis Ther. 2015;3:8-20.

  19. Wong KH, Skelton SK. New, practical approach to detecting antibody to pertussis toxin for public health and clinical laboratories. J Clin Microbiol. 1988;26(7):1316-20.

  20. Centro para el Control Estatal de la Calidad de los Medicamentos (CECMED). Regulación No. 41-2007: Validación de Métodos Analíticos. Ámbito Regulador. 2007;(53):1-14.

  21. International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use. Validation of Analytical Procedures: Text and Methodology Q2 (R1). ICH; 2005 Nov.

  22. Cembrowski GS, Sullivan AM. Quality control and statistics. In: Bishop ML, Duben-Engelkirk JL, Fody EP, eds. An Introduction to Clinical Chemistry. Philadelphia, USA: Lippincott. p. 63-101.

  23. Thalen M, van der Ark A, van den Ijssel J, van Straaten I, Jansen D, Beuvery C, et al. Improving the cellular pertussis vaccine: increased potency and consistency. Vaccine. 2008;26(5):653-63.

  24. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227 (5259):680-5.

  25. Fernandez-Patron C, Castellanos-Serra L, Rodriguez P. Reverse staining of sodium dodecyl sulfate polyacrylamide gels by imidazole-zinc salts: sensitive detection of unmodified proteins. Biotechniques. 1992;12(4):564-73.

  26. Layne E. Spectrophotometric and turbidimetric methods for measuring proteins. Methods Enzymol. 1957;3:447–54.

  27. Welfringer F, d’Athis P, Scherrmann JM, Herve F. Development and validation of an antigen-binding capture ELISA for native and putrescine-modified anti-tetanus F(ab’)2 fragments for the assessment of the cellular uptake and plasma kinetics of the antibodies. J Immunol Methods. 2005;307(1-2):82-95.

  28. Findlay JW, Smith WC, Lee JW, Nordblom GD, Das I, DeSilva BS, et al. Validation of immunoassays for bioanalysis: a pharmaceutical industry perspective. J Pharm Biomed Anal. 2000;21(6):1249-73.

  29. Riley CM, Rosanke TW. Development of validation of analytical methods: progress in pharmaceutical and biomedical analysis. vol 3. New York: Elsevier (Pergamon), NY; 1996.

  30. Shah VP, Midha KK, Dighe S, McGilveray IJ, Skelly JP, Yacobi A, et al. Analytical methods validation: bioavailability, bioequivalence and pharmacokinetic studies. Conference report. Eur J Drug Metab Pharmacokinet. 1991;16(4):249-55.

  31. ICH. Guideline on validation of analytical procedures: definitions and terminology International Conference of Harmonization (ICH) of Technical Requirements for the Registration of Pharmaceuticals for Human Use. Geneva: ICH; 1996.

  32. Cummings J, Ward TH, Greystoke A, Ranson M, Dive C. Biomarker method validation in anticancer drug development. Br J Pharmacol. 2008;153(4):646-56.

  33. Guidance for Industry: Bioanalytical. Method Validation. Rockville, MD: US Department of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research; 2001.

  34. DeSilva B, Smith W, Weiner R, Kelley M, Smolec J, Lee B, et al. Recommendations for the bioanalytical method validation of ligandbinding assays to support pharmacokinetic assessments of macromolecules. Pharm Res. 2003;20(11):1885-900.

  35. Cembrowski GS, Sullivan AM. Quality control and statistics. In: An Introduction to Clinical Chemistry, Bishop ML, Duben-Engelkirk JL, Fody EP, eds. Philadelphia, USA: Lippincott. p. 63-101.

  36. Fuge J. FDA CONFERENCE Report: Validation considerations when preparing for FDA inspections. J Validat Technol. 2005;11(3):253-6.




2020     |     www.medigraphic.com