Development and Validation of Enzyme-linked Immunosorbent Assay for Anti-mouse Pertussis Immunoglobulin G Using International Reference Anti- Mouse Serum NIBSC 97/642

Overview

Journal Clin Exp Vaccine Res

Date 2024 Aug 15

PMID 39144122

Authors

Kyu-Ri Kang

Yi-Hyeon Kwon

Gyu-Won Cho

Gi-Sub Choi

Joon-Hwan Ji

Hyun-Mi Kang

Soo-Young Lee

Jin-Han Kang

Affiliations

Soon will be listed here.

Abstract

Purpose: In this study, an in-house enzyme-linked immunosorbent assay (ELISA) was developed and validated. The titer of ELISA was calculated using the reference line (RFL) method based on the standard curve drawn using the international reference anti-mouse serum NIBSC (National Institute for Biological Standards and Control) 97/642.

Materials And Methods: In the development step, signal to noise was depicted to select the buffers that showed the most appropriate ratio. In the validation step, standard range, precision, dilution linearity, and specificity were confirmed, and RFL and parallel line (PLL) methods were compared in precision and dilution linearity.

Results: Coating concentration for plate was achieved at 0.1 µg/mL for pertussis toxin (PT), 0.15 µg/mL for filamentous hemagglutinin antigen (FHA), and 0.25 µg/mL for pertactin (PRN). The signal to noise ratio was 22.02 for PT, 14.93 for FHA, and 8.02 for PRN with 0.25% goat serum in phosphate-buffered saline (PBS) as a dilution buffer, and 2% skim milk in PBS as a blocking buffer. Based on the precision results, we assessed the lower limit of quantification by 1, 0.2, and 1.5 EU/mL concentration for PT, FHA, and PRN which met the ICH (International Council for Harmonization) M10 criteria of a 25% accuracy and total error of 40%. In specificity, homologous serum was spiked into heterologous serum and the accuracy met the criteria. There was no difference in the results between RFL and PLL calculations (p-value=0.3207 for PT, 0.7394 for FHA, 0.2109 for PRN).

Conclusion: ELISA validated with RFL calculation method in this study is a relatively accurate assay for mouse humoral immunogenicity test.

References

Sato H, Sato Y . Bordetella pertussis infection in mice: correlation of specific antibodies against two antigens, pertussis toxin, and filamentous hemagglutinin with mouse protectivity in an intracerebral or aerosol challenge system. Infect Immun. 1984; 46(2):415-21. PMC: 261548. DOI: 10.1128/iai.46.2.415-421.1984. View

Das R, Xing D, Rigsby P, Newland P, Corbel M . International collaborative study: evaluation of proposed International Reference Reagent of pertussis antiserum (mouse) 97/642. Biologicals. 2001; 29(2):137-48. DOI: 10.1006/biol.2001.0288. View

Coulson B, Grimwood K, Bishop R, Barnes G . Evaluation of end-point titration, single dilution and capture enzyme immunoassays for measurement of antirotaviral IgA and IgM in infantile secretions and serum. J Virol Methods. 1989; 26(1):53-65. DOI: 10.1016/0166-0934(89)90074-8. View

Mills K, Ryan M, Ryan E, Mahon B . A murine model in which protection correlates with pertussis vaccine efficacy in children reveals complementary roles for humoral and cell-mediated immunity in protection against Bordetella pertussis. Infect Immun. 1998; 66(2):594-602. PMC: 107945. DOI: 10.1128/IAI.66.2.594-602.1998. View

Andre M, Poirier B, Bornstein N, Marmonier D, El Zaouk A, Fuchs F . Key points for the development of mouse immunogenicity test as potency assay for acellular pertussis vaccines. Biologicals. 2001; 28(4):217-25. DOI: 10.1006/biol.2000.0259. View

Libster R, Edwards K . Re-emergence of pertussis: what are the solutions?. Expert Rev Vaccines. 2012; 11(11):1331-46. DOI: 10.1586/erv.12.118. View

Kadam L, Patel K, Gautam M, Thorat S, Kale P, Ghule A . Development and validation of magnetic bead pentaplex immunoassay for simultaneous quantification of murine serum IgG antibodies to acellular pertussis, diphtheria and tetanus antigens used in combination vaccines. Methods. 2019; 158:33-43. DOI: 10.1016/j.ymeth.2019.01.015. View

Chiappini E, Stival A, Galli L, de Martino M . Pertussis re-emergence in the post-vaccination era. BMC Infect Dis. 2013; 13:151. PMC: 3623740. DOI: 10.1186/1471-2334-13-151. View

Watanabe M, Funaishi K, Takeo T, Endoh M . Efficacy of pertussis vaccines consisted of antigens detoxified with tea-leaf catechins. Vaccine. 2001; 19(9-10):1204-10. DOI: 10.1016/s0264-410x(00)00307-8. View

10.

Peterman J, Butler J . Application of theoretical considerations to the analysis of ELISA data. Biotechniques. 1989; 7(6):608-15. View

11.

Choi G, Huh D, Han S, Ahn D, Kang K, Kim J . Enzyme-linked immunosorbent assay for detecting anti-pertussis toxin antibody in mouse. Clin Exp Vaccine Res. 2019; 8(1):64-69. PMC: 6369130. DOI: 10.7774/cevr.2019.8.1.64. View

12.

Stenger R, Smits M, Kuipers B, Kessen S, Boog C, van Els C . Fast, antigen-saving multiplex immunoassay to determine levels and avidity of mouse serum antibodies to pertussis, diphtheria, and tetanus antigens. Clin Vaccine Immunol. 2011; 18(4):595-603. PMC: 3122557. DOI: 10.1128/CVI.00061-10. View

13.

Cheung G, Xing D, Prior S, Corbel M, Parton R, Coote J . Effect of different forms of adenylate cyclase toxin of Bordetella pertussis on protection afforded by an acellular pertussis vaccine in a murine model. Infect Immun. 2006; 74(12):6797-805. PMC: 1698075. DOI: 10.1128/IAI.01104-06. View

14.

Grabowska K, Wang X, Jacobsson A, Dillner J . Evaluation of cost-precision rations of different strategies for ELISA measurement of serum antibody levels. J Immunol Methods. 2002; 271(1-2):1-15. DOI: 10.1016/s0022-1759(02)00334-4. View

15.

Sato Y, Sato H . Development of acellular pertussis vaccines. Biologicals. 1999; 27(2):61-9. DOI: 10.1006/biol.1999.0181. View

16.

Watanabe M, Nagai M . Reciprocal protective immunity against Bordetella pertussis and Bordetella parapertussis in a murine model of respiratory infection. Infect Immun. 2001; 69(11):6981-6. PMC: 100078. DOI: 10.1128/IAI.69.11.6981-6986.2001. View

17.

Plikaytis B, Turner S, Gheesling L, Carlone G . Comparisons of standard curve-fitting methods to quantitate Neisseria meningitidis group A polysaccharide antibody levels by enzyme-linked immunosorbent assay. J Clin Microbiol. 1991; 29(7):1439-46. PMC: 270131. DOI: 10.1128/jcm.29.7.1439-1446.1991. View

18.

Althouse B, Scarpino S . Asymptomatic transmission and the resurgence of Bordetella pertussis. BMC Med. 2015; 13:146. PMC: 4482312. DOI: 10.1186/s12916-015-0382-8. View

19.

Reizenstein E, Hallander H, Blackwelder W, Kuhn I, Ljungman M, Mollby R . Comparison of five calculation modes for antibody ELISA procedures using pertussis serology as a model. J Immunol Methods. 1995; 183(2):279-90. DOI: 10.1016/0022-1759(95)00067-k. View

20.

Cherry J . Epidemic pertussis in 2012--the resurgence of a vaccine-preventable disease. N Engl J Med. 2012; 367(9):785-7. DOI: 10.1056/NEJMp1209051. View