Development of a New Approach to Determine the Potency of Bacille Calmette–Guérin Vaccines Using Flow Cytometry.
10.24171/j.phrp.2017.8.6.06
- Author:
Eunjeong GWEON
1
;
Chanwoong CHOI
;
Jaeok KIM
;
Byungkuk KIM
;
Hyunkyung KANG
;
Taejun PARK
;
Sangja BAN
;
Minseok BAE
;
Sangjin PARK
;
Jayoung JEONG
Author Information
1. Biologics Research Division, National Institute of Food and Drug Safety Evaluation, Cheongju, Korea. 0jjy@korea.kr
- Publication Type:Original Article
- Keywords:
BCG vaccine;
vaccine potency;
flow cytometry
- MeSH:
Administration, Cutaneous;
BCG Vaccine;
Cell Count;
Flow Cytometry*;
Fluorescein;
Methods;
Mycobacterium bovis;
Quality Control;
Stem Cells;
Vaccine Potency;
Vaccines*
- From:
Osong Public Health and Research Perspectives
2017;8(6):389-396
- CountryRepublic of Korea
- Language:English
-
Abstract:
OBJECTIVES: To circumvent the limitations of the current golden standard method, colony-forming unit (CFU) assay, for viability of Bacille Calmette–Guérin (BCG) vaccines, we developed a new method to rapidly and accurately determine the potency of BCG vaccines. METHODS: Based on flow cytometry (FACS) and fluorescein diacetate (FDA) as the most appropriate fluorescent staining reagent, 17 lots of BCG vaccines for percutaneous administration and 5 lots of BCG vaccines for intradermal administration were analyzed in this study. The percentage of viable cells measured by flow cytometry along with the total number of organisms in BCG vaccines, as determined on a cell counter, was used to quantify the number of viable cells. RESULTS: Pearson correlation coefficients of FACS and CFU assays for percutaneous and intradermal BCG vaccines were 0.6962 and 0.7428, respectively, indicating a high correlation. The coefficient of variation value of the FACS assay was less than 7%, which was 11 times lower than that of the CFU assay. CONCLUSION: This study contributes to the evaluation of new potency test method for FACS-based determination of viable cells in BCG vaccines. Accordingly, quality control of BCG vaccines can be significantly improved.
