earticle

영어

Quantitative measurement of anti-HBs is used to evaluate the response to hepatitis B vaccination in health care workers and also used to potency test for several immunoglobulin preparations as an effectiveness factor. Comparison of 5 assay methods for anti-HBs is performed to improve potency assay of Human Immunoglobulin G(HuIgG) preparation. This study established EIA method for anti-HBs comparing with current MEIA method. Three commercial EIA kits were compared with co-laboratory studies including the HuIgG manufacturers and Korea FDA using national standard (NS) and international standard (IS) of anti-HBs. Resulted similar reproducibility (average CV=5%, recovery >96%) of each kits however, Enzygnost was selected as an optimal EIA kit which showed the most stable precision (CV=4.1%) when the negative plasma was applied as diluent for sample and standard. Even though negative plasma and 5% human serum albumin (HSA) showed compatible reproducibility, negative plasma gave better precision (CV=4.1% vs 7.1%) and lower background. Using the optimal EIA kit, validation were performed including intra- and inter-day assays, inter-laboratory assay and inter-method comparison with 22 representative HuIgG preparations. The samples consist of 9 lot of maltose added HuIgG, 3 lot of HuIgG, 5 lot of Anti-HBV HuIgG and 5 NS spiked samples (total 22 samples). As a result intra day precision was 4.6±2.0% and inter day precision was 7.7±5.3 for EIA (N=180 for 5 days), 3.7±1.9 for AxSym (N=10 for 5 days) and 1.6±0.8 for IMx(N=6 for 2 days). As an accurate test with the results of NS spiked samples, the mean recovery was 100.1±12.6%. The statistical analysis showed that the detection limit of EIA was below 10 IU/L and the range was 10~150 IU/L. The linearity of dose response curve with NS were confirmed (R2>0.998) and robustness was proved based on the spiking assay with representative samples (100.1%) which showed no matrix effect. Therefore, the EIA assay was validated and showed relevance to MEIA including two automated instruments; IMx and AxSYM of Abbott. As a conclusion the EIA assay could replace the current MEIA method.