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Indirect ELISA Method for Measurement of Lactoperoxidase using IgY Antibody  

이승배 (상지대학교 생명공학과)
최석호 (상지대학교 생명공학과)
최재원 (상지대학교 생명공학과)
Publication Information
Food Science of Animal Resources / v.24, no.2, 2004 , pp. 182-188 More about this Journal
Abstract
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 ${\alpha}$-lactalbumin, ${\beta}$-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 ${\alpha}$-lactalbumin, ${\beta}$-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 0.25$\mu\textrm{g}$/mL and 1:8,000 respectively. Sensitivity in the standard curve of LPO was ranged from 0.01 to 1$\mu\textrm{g}$/mL using anti-LPO IgY.
Keywords
lactoperoxidase; anti-lactoperoxidase IgY; indriect ELISA;
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