摘要
【目的】探索产肠毒素大肠杆菌(ETEC)987P菌株培养的最佳条件及987P菌毛诱发蛋鸡制备卵黄抗体(IgY)的最佳免疫程序,为IgY的工业化、规模化生产提供参考。【方法】扩大培养987P菌株,抽提层析纯化987P菌毛蛋白,然后以其为抗原免疫20周龄开产海兰蛋鸡,采用间接ELISA检测987P菌毛蛋白免疫IgY效价。【结果】987P菌株的最佳培养条件为37℃下Slanctz固体培养基培养24h;以纯化987P菌毛蛋白为抗原免疫20周龄开产蛋鸡,发现987P菌毛蛋白二免、三免IgY效价总体趋势基本一致,均在第3周达到峰值并维持1周,二免抗体峰值为213,三免抗体峰值是二免峰值的2倍(214),但第4周过后三免IgY效价仍保持在213,且持续时间长,一般维持2～3周。【结论】以纯化987P菌毛蛋白为抗原免疫蛋鸡,能获得较高效价的IgY,且适当增加免疫次数能有效提高抗体水平和延长持续时间。
[Objective]The present experiment was aimed to develop the best culture conditions for enterotoxigenic Escherichia coli（ETEC） 987P and the best immunity procedure for preparation of yolk antibody（IgY） with layer induced by 987P pili to provide reference for industrial scale production of IgY.[Method]The strain 987P was cultured and its pili protein was extracted and purified.The purified pili protein was used as antigen to immune 20-week egg-production Hailan layer,and indirect ELIZA method was used for detection of IgY titer of pili protein immunity yolk antibody.[Result]The result showed that the best culture conditions of 987P was in Slanctz solid culture medium for 24 hours at 37℃.Using purified pili protein as antibody to immune 20-week layer,the IgY titers of 2 n d and 3 r d immunity in 987P pili protein showed similarity change trends,which attained the peak and kept constant for a week.The antibody titer peak of 2 n d and 3 r d immunity was 213 and 214.After 4 t h week,the IgY titer of 3 r d immunity still kept at 213 for a long time of 2-3 weeks.[Conclusion]The higher titer IgY may be obtained by using purified 987P pili as antibody to immune layer,and the antibody level can be enhanced and its duration can be elongated with suitably increased immunity times.
出处
《南方农业学报》
CAS
CSCD
2011年第10期1284-1287,共4页
Journal of Southern Agriculture
基金
广西大学研究生创新计划项目(2007105930905M079)
