腐败希瓦氏菌卵黄抗体活性片段的制备及抑菌效果的分析
详细信息Production and antibacterial activity evaluation offragments from chicken egg yolk Immunoglobulin against Shewanellaputrefaciens
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摘要: 为探讨卵黄抗体活性片段的制备方法,利用胃蛋白酶对腐败希瓦氏菌卵黄抗体进行酶解,通过高效液相色谱法动态分析酶解过程,建立了一套有效的纯化方法,并对活性片段抑菌效果进行鉴定分析。结果表明,经胃蛋白酶作用6h后,卵黄抗体被酶解为43000u左右及其他更小分子量的蛋白片段,且组分不再随时间发生变化。酶解混合液经Sephacryl S-100层析纯化后,即得纯度大于90%的活性蛋白片段。此蛋白片段保留了较高的抗原结合能力,且抑菌效果明显。研究为制备高纯度的卵黄抗体活性片段提供有效方法,为进一步扩大腐败希瓦氏菌卵黄抗体应用范围奠定理论基础。Abstract: Methods were described for the production and purification of active fragments from chicken egg yolk Immunoglobulin (IgY) against Shewanellaputrefaciens. The peptic digestion procedure monitored by high performance liquid chromatograph (HPLC) , and the antibacterial activity of the fragments was also analyzed. Results demonstrated that IgY was divided into protein fragments of 43000u and other small ones by incubating with pepsin for 6h, and after 6h, the components of the fragments did not change any more.Active protein fragment of 90% purity was obtained using Sephacryl S-100 gel filtration chromatography and it retained antigenbinding and antibacterial activities.This research provided an effective method to produce active fragments from IgY, laying a theoretical foundation for expanding the application of IgY against Shewanellaputrefaciens.
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