Clin Exp Vaccine Res.  2019 Jan;8(1):35-42. 10.7774/cevr.2019.8.1.35.

Development and implementation of standardized method for detecting immunogenicity of acellular pertussis vaccines in Korea

• Park C
• Huh DH
• Han SB
• Choi GS
• Kang KR
• Kim JA
• Kang JH

• KMID: 2438960
• DOI: http://doi.org/10.7774/cevr.2019.8.1.35

Abstract

PURPOSE
There is no standard method for confirming the immunogenicity of acellular pertussis vaccines. We tried to develop a local standard method for evaluating the immunogenicity of the three-component of acellular pertussis vaccines which was developed by a Korean local company.
MATERIALS AND METHODS
The developed pertussis antigens (pertussis toxin, filamentous hemagglutinin, pertactin) were evaluated by in-house enzyme-linked immunosorbent assay (ELISA) using 189 negative sera, 25 positive sera, and 73 paired sera (pre- and post-Tdap [tetanus, diphtheria, and acellular pertussis] vaccinated sera). ELISA units were calculated by the reference line method, compared with World Health Organization reference sera, and the cut-off value was calculated using negative sera.
RESULTS
When compared to National Institute for Biological Standards and Control control antigen (NIBSC) control antigens, the developed pertussis toxin (PT) and filamentous haemagglutinin (FHA) antigens were 203.48 and 61.60 IU/µg, respectively. Each in-house ELISA was established by validating the coefficients of variation % (PT, 11.53%; FHA, 8.60%; pertactin [PRN], 9.86%) obtained from the results of inter- and intra-assay variation. Also, the cut-off values of PT, FHA, and PRN were 11.65, 38.95, and 5.66 EU/mL, respectively. The distributions of antibody levels in paired showed that 93.15% (68/73) in anti-PT IgG, 97.26% (72/73) in anti-FHA IgG, and 100% in anti-PRN IgG were higher than a 100% increase after vaccination. Additionally, the values of 89.04% (65/73) in anti-PT IgG, 97.26% (72/73) in anti-FHA IgG, and 100% in anti-PRN IgG were below each cut-off point.
CONCLUSION
We established an in-house ELISA method using self-developed antigens, and these immunoassays have provided a way to standardize measuring the immunogenicity of newly developed vaccines, through single- and dual-serology.

Keyword

Pertussis vaccine; Pertussis toxin; FHA protein; Pertactin; Enzyme-linked immunosorbent assay; Serology

MeSH Terms

Diphtheria
Enzyme-Linked Immunosorbent Assay
Hemagglutinins
Immunoassay
Immunoglobulin G
Korea*
Methods*
Pertussis Toxin
Pertussis Vaccine
Vaccination
Vaccines*
Whooping Cough*
World Health Organization
Hemagglutinins
Immunoglobulin G
Pertussis Toxin
Pertussis Vaccine
Vaccines

Figure

• 

  Fig. 1 Analysis of anti-pertussis antigens (A, PT; B, FHA; C, PRN), IgG in the pre- and post-vaccinated sera group, and increased ratio between A and B Tdap vaccination group. PT, perussis toxin; FHA, filamentous haemagglutinin; PRN, pertactin; Tdap, tetanus, diphtheria, and acellular pertussis.

• 

  Fig. 2 Results of dual-serology (paired pre- and post-vaccinated sera) by established in-house enzyme-linked immunosorbent assay. X-axis indicates paired sample, while y-axis indicates anti-PT IgG (A), anti-FHA IgG (A), and anti-PRN IgG (C), respectively. Black squares indicate post-vaccinated sera, while blank circles indicate pre-vaccinated sera. PT, perussis toxin; FHA, filamentous haemagglutinin; PRN, pertactin

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