Determination of Analytical Method Validation for Indirect ELISA Quantification of Antibody Titer against PT Antigen in Acellular Pertussis Vaccine Using Mouse Immunogenicity Test

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Apichai Supasansatorn Supasansatorn
Ratchanon Rakcheep
Assjan Amen
Wereyamarst Jaroenkunathum
Supaporn Phumiamorn

Abstract

Acellular pertussis vaccines are contained pertussis toxin (PT) as a major active substrate
component, which are produced in Thailand. Then, the researchers developed a method for
determination of the anti-PT antibody titer used to control the potency of the vaccine using
mouse immunogenicity test (MIT). The MIT is a non-lethal animal model designed to evaluate
antibody responses in immunized mice to the PT antigen claimed to contribute to vaccine
efficacy by ELISA technique. The validation method showed a good specification by separation
of anti-PT antibody titer between vaccine test group and negative control group. The linearity
was found at the range of anti-PT antibody titer from 1.95 to 4000.00 mIU/mL, with the coefficient
of correlation and the coefficient of determination were greater than 0.99. The lower limit of
quantitation was 1.95 mIU/mL and the upper limit of quantitation was 4,000.00 mIU/mL. The
range ranged from 7.81 to 500.00 mIU/mL. The accuracy had the percentage relative error (%RE)
between -2.11 to 2.37%. The precision varied from 5.57 to 11.51 percentage of variance
coefficient (%CV). The method was robust to the test system and analyzers, %CV was 6.47 to
9.08. Based on the validation results, the method demonstrates that the method is reliable
and suitable. It can be used as a standard method for the detec tion of anti-PT antibody to
effectively control the quality of acellular pertussis vaccine.

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References

1. สำนักโรคติดต่อทั่วไป กรมควบคุมโรค กระทรวงสาธารณสุข. ตำราวัคซีนและการสร้างเสริมภูมิคุ้มกันโรคปี 2556. พิมพ์ครั้งที่ 1. นนทบุรี:มหาวิทยาลัยราชภัฎสวนสุนันทา;2556

2. Stanley A. Plotkin, Water A. Orenstein and Paul A. Offit. Vaccine 6 TH Edition,Philadelphia, PA: Elsevier, 2013: 449-50

3. WHO Expert Committee on Biological Standardization. WHO Technical Report Series No. 979; 2013: 205-206, 241-4.

4. European Directorate for the Quality of Medicines and HealthCare. European pharmacopoeia 8.0: Assay of pertussis vaccine (acellular). Nördlingen: Druckerei;2014: 252-5.

5. International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use. Validation of analytical procedures: text and methodology Q2 (R1). Geneva: ICH Steering Committee;Nov 2005

6. R.G. Das, D. Xing, P. Newland and MJ Corbel. International collaborative Study: Evaluation of Proposed International Reference Reagent of Pertussis Antiserum Mouse) 97/642 Biologicals (2001) 29,137-48.