Validation of Analytical Method for Hepatitis B Component in Pentavalent Vaccine as Well as Hepatitis B Vaccine (Monovalent) by Sandwich Elisa Method

For evaluating the antigen content in recombinant Hepatitis B Vaccine (Monovalent and Pentavalent vaccines), National Control Laboratory, Kasauli has taken an initiative to develop a uniform method for evaluation of antigen content of Hepatitis B vaccine. All the manufacturers in India are having different method for evaluating Relative Potency of Hepatitis B Vaccine. In the present study the samples of different manufacturers were tested for potency by Sandwich ELISA method. This method was validated to check the suitability for all the manufacturers. Various validation parameters were evaluated to verify the fulfilled defined acceptance criteria. This validation method of Potency estimation can be transferred to different manufacturers throughout India to maintain the synchronization and harmonization of potency estimation of Hepatitis B vaccine. For the invitro potency assay of Hepatitis B Vaccine Adaltis EIAgen HBsAg Kit was used to determine the Relative Potency. Validation Parameters were evaluated following the International Conference on Harmonization (ICH) guidelines. Samples of different Manufacturers were tested for validation parameters. It was confirmed that the ELISA Kit when used for vaccine potency testing meets the criteria of specificity , accuracy, linearity, precision (Intra assay precision(repeatability), Inter assay precision(reproducibility & ruggedness).The method was found specific based on the results generated by ELISA Kit, when used for potency testing , meets the criteria for accuracy (80-125%), linearity R ≥ 0.99, precision (repeatability , with a CV% less than 15% and reproducibility with a CV% less than 15%).