Validation study of a quantitative ELISA assay to determine the potency of human immunoglobulin containing antibody to hepatitis B surface antigen

Abstract

Background/Purpose: The quantitative ELISA method is a fast and accurate method. The
advantages of this method include long shelf life of test kit, simple procedure and cost
effectiveness. This study was conducted to validate the application of a quantitative
ELISA assay and determine if this technique is suitable for quality control of human
immunoglobulin products containing antibody to hepatitis B surface antigen.
Methods: descriptive laboratory study. We explored the technical specifications of a
commercial quantitative ELISA assay for the determination of anti-HBs antibodies. They
included linearity, limit of quantitation, trueness, precision and robustness of the ELISA
kit. This commercial quantitative ELISA kit has been approved by the European
Regulatory Authority.
Results: Our results included as follow: for the linearity of the ELISA assay, the correlation
coefficient (R) was 0.99 with CV 1.57%, showing a strong realtionship between anti-HBs
concentration and OD; limit of quantitation (LoQ) was 36,68 (IU/ml), CV of intra-run and
inter-run precision were 3.98% and 4-6%, respectively. Results of trueness and robustness
showed t < tα. All of the technical specifications of the ELISA assay were passed for
quality requirements and suitable for use to determine the potency of the ImmunoHBs
180IU/ml product.
Conclusion: Quantitative ELISA method is a suitable immuno method to determine the
titer of anti-HBs in ImmunoHBs 180IU/ml and can be used as a potency test for quality
control of human immunoglobulin containing antibody to hepatitis B surface antigen.