猪圆环病毒2型重组Cap蛋白体外稳定性研究

Study on Stability of Capsid Protein of Porcine Circovirus Type 2 in vitro

为筛选猪圆环病毒2型(PCV2)Cap蛋白在体外的稳定性条件,采用建立的PCV2双抗夹心ELISA方法,对Cap蛋白在不同贮藏条件下的抗原活性进行了检测。结果表明,Cap蛋白在25℃以下贮存时,其活性保持稳定;贮存时温度变化对蛋白质稳定性影响较大;在甘氨酸和甘油2种蛋白保护剂存在条件下,蛋白质活性稳定性增强。表明,Cap蛋白的体外贮存、改造及病毒样颗粒组装应在25℃以下进行,尽量避免温度剧烈变化,同时添加蛋白质保护剂有利于蛋白质的体外稳定。

The present study was aimed to obtain the optimal storage of the Cap protein of porcine circo-virus type 2 （ PCV2 ） . Double antibody sandwich ELISA method was established to determine the bioacti-vity of Cap protein treated under different storage conditions. The results showed that the bioactivity of Cap protein remained stable when storaged under 25 ℃,and temperature shifts would have a significant impact on the bioactivity of Cap protein. Protein stabilizer helped remaining the immunoreactivity of Cap protein. Accordingly,further modification or assembly of Cap protein into virus-like particles should be performed under 25 ℃, and avoid rapid changes in temperature with the addition of protein stabilizer.