봉독의 항독소(IgY)생산을 위한 실험적 연구

Abstract

This study was carried out for production of neutral antibody to bee venom . Hen layings were injected repeatedly with bee venom and phospholipase with Freund`s adjuvant. Specific antibody in egg yolk from immunized hen laying was separated, and purified, also immunological characteristics of anti phospholipase was invested. The results were summarized as follows: 1. Phospholipase was showed single band at molecular weight 17,000 in SDS-PAGE and bee venom was showed two band at molecular weight 17,000 and under molecular weight 6,500 in SDS-PAGE. 2. During 70 days after hen immunized with bee venom and phospholipase , antibodies(anti-bee venom IgY) to bee venom were showed poor ELISA value in egg yolk, but antibodies to phospholipase in egg yolk were increased ELISA value from 8 days or 15 days and found maximum ELISA value at 42 days. Also after booster at 49 days, ELISA value of anti Phospholipase in egg yolk was supported at optical density(O.D) 1.0 level, continuously. 3. Titer of phospholipase was showed 1: 32,000. 4. In double immunodiffusion test to phospholipase after double dilution of anti-phospholipase , only precipitation line was made in 1:1 dilution well of anti-Phospholipase . But In immunodiffusion test to anti-phospholipase after double dilution of phospholipase , Precipitation line to 250ul/ml well of phospholipase was showed. In double immunodiffusion test to bee venom(1mg/ml) after double dilution anti-phospholipase , all well without 1:32 dilution well were showed strong precipitation line. 5. In dot bloting test to anti-phospholipase after diluting bee venom(0.5mg/ml), dot bloting color was showed clearly to in bee venom.