口蹄疫病毒部分免疫功能肽段的合成、鉴定及免疫功能评价

Characterization and evaluation of immune function on synthesized FMDV peptides

利用微波多肽合成仪Liberty,合成O型口蹄疫病毒VP1 140～160和200～213氨基酸序列的两条肽段,并用高压液相色谱仪及质谱分析仪就合成的多肽片段进行分析纯化和鉴定;将合成的多肽片段作为半抗原与载体蛋白BSA偶联,免疫小鼠,进行血清制备和抗体效价测定.结果表明：成功合成了口蹄疫2条免疫功能短肽,其纯度均达到80%以上.半抗原成功与载体蛋白偶联;小鼠抗血清经过抗体效价检测证明半抗原与载体蛋白偶联组的抗体水平高于其余2个对照组,但是达不到免疫保护的水平.同时说明Liberty是一种操作简单、方便,合成效率高的多肽合成仪,可高效合成研究用多肽.

Type O FMDV VP1 140～160and 200～213amino acid fragments were synthesized by LibertyTM automatic microwave peptide synthesizer.Two polypeptides were purified and identified by HPLC and MS.Mice were immunized by polypeptide conjugated to BSA and antibody titers were measured.The results showed that the two peptides were successfully synthesized and their purity was more than 80%.As the haptens,they were effectively conjugated with the carrier protein.Their anti-mouse serum antibody titers were higher than that of the other two negative control groups;however the antibody levels of immune animals were not up to the scope of protection.It also proved that LibertyTM was a simple,convenient and efficient peptide synthesizer which could synthesis research-grade peptides efficiently.This study supported evidence and a new method of antigen peptide synthesis to prepare for vaccine research.