IgY 항체를 이용하여 Lactoperoxidase 정량을 측정하기 위한 Indirect ELISA 방법의 개발

To determine the concentration of lactoperoxidase (LPO), an indirect enzyme-linked immunosorbant assay (ELISA) was developed. Anti-LPO egg yolk immunoglobulin (IgY) was transferred to egg yolk by immunizing of Brown hens with LPO. The titer of purified anti-LPO IgY was 1: 520,000. The immunological response of anti- LPO IgY with α-lactalbumin, β-lactoglobulin, casein and lysozyme were evaluated, resulting that the anti-LPO IgY found to be a specific antibody toward LPO and no cross-reaction was observed against α-lactalbumin, β-lactoglobulin, casein, and lysozyme in double immunodiffusion test and ELISA test. In indirect ELISA method, coating concentration of LPO and dilution rate of anti-LPO IgY was 0.25ug/mL and 1:8,000 respectively. Sensitivity in the standard curve of LPO was ranged from 0.01 to lug/mL using anti-LPO IgY.