ABSTRACT
Quantification of hepatitis B surface antigen (HBsAg) or relative
in vitro
potency in the final vaccines is a prerequisite for hepatitis B vaccine batch release. The commercial kit for ...automated analysis (AxSYM) is expensive, and an alternative is required for the estimation of HBsAg in hepatitis B vaccines. Mouse monoclonal antibodies (MAbs) specific for HBsAg were developed and characterized. One of the monoclonal antibodies (HBs06) was used in development of an immunocapture ELISA (IC-ELISA) as an unlabeled capture antibody and biotin-labeled detection antibody. The IC-ELISA was standardized and validated using experimental hepatitis B vaccine batches with various HBsAg concentrations per dose and commercial vaccines. The vaccine was treated with an alkaline solubilizer to desorb the HBsAg from Algel-adjuvanted vaccines before testing, and the sensitivity of the test was 5 ng/ml. A good correlation could be observed between the HBsAg estimates derived by both formats, except for the higher HBsAg concentration range, where the IC-ELISA format could estimate closer to the actual values than AxSYM. There was a significant correlation between the estimated relative potencies of the two methods. There was lack of correlation between the
in vivo
potency and the relative
in vitro
potency. However, the estimates of IC-ELISA were comparable to the
in vivo
values when compared with the estimates of AxSYM. The IC-ELISA can therefore be considered to be a reliable test for deriving
in vitro
relative potency and antigen concentration in vaccine batches for batch control and release.
Laboratory detection of specific foot-and-mouth disease virus (FMDV) is routinely carried out by ELISA and RT-PCR. Identification and serotyping of FMDV by ELISA requires polyclonal antibodies raised ...in rabbits and guinea pigs. The polyclonal antibodies have certain disadvantages such as batch to batch variation, inconsistent yields of antibodies and limited quantity of serum obtained from individual animals. This paper describes a method wherein monoclonal antibodies and chicken IgY were used in an antigen capture-ELISA for serotyping of thirty tongue epithelial samples and sixty tissue culture fluids. The results were compared with the routine antigen detection ELISA. The present study indicated that monoclonal antibodies and chicken IgY can substitute conventional polyclonal antibodies for routine serotyping of FMDV.
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