摘要
目的:用人脂蛋白脂酶制备鸡卵黄抗体,并研究其应答持久性、稳定性.方法:用纯化的人脂蛋白脂酶免疫生蛋鸡,琼脂糖免疫扩散法测定IgY抗体效价变化.PEG法结合Sephadex-150凝胶过柱提取抗体;ELISA法测抗体对酸、碱、酶的稳定.结果:免疫后大约3～4周抗体效价达到高值,此高效价可持续3个月;抗体含量为8.11mg/ml蛋黄;IgY在70℃以下基本不失活;在pH4～10时活性无明显变化;胃蛋白酶处理90min后IgY活性稳定.结论:用人脂蛋白脂酶免疫鸡可制备高效价的特异抗体IgY,抗体应答时间持久、稳定性好.
Objective: To investigate the response-persistence and stability of egg yolk antibody(IgY)of anti-lipoprotein lipase.Method:Immunize hen with human lipoprotein lipase.The titer of IgY was measured by agarose immunodiffusion assay.The IgY was extracted from the egg yolk by PEG and further purified by Sephadex-150 gel chromatography.The stability of IgY against pH and enzyme was detected by enzyme labeling linked immunosorbent assay(ELISA).Results:The titer of antibody raised gradually within 2 weeks after the first immunization,and reached at the peak within 3～4 weeks.This high titer could maintain for 3 months.8.11mg IgY could be obtained from 1 ml egg yolk.The IgY was very stable at the condition of <70℃,pH4～10,and its activity was still preserved after 90 minutes treatment of pepsin solution.Conclusion:The specific IgY of anti-lipoprotein lipase has the characteristics of high titer,stability and long response-persistence.
出处
《浙江中医学院学报》
2005年第3期51-52,55,共3页
Journal of Zhejiang College of Traditional Chinese Medicine
基金
国家自然科学基金课题资助
批准号30070932
