摘要
目的设计合成PCV2 CAP的B细胞表位抗原肽并对其进行鉴定。方法采用DNAStar和BcePred分析软件并结合吴玉章氨基酸抗原指数计算方法预测PCV2 CAP的B细胞表位．以此合成4分支多重抗原肽结构．同时串联一个通用型n表位；采用Fmoc法合成4分支PCV2CAP多重抗原肽（MAP），通过高效反相液相色谱（RP-HPLC）、质谱（MS）分析对其进行初步鉴定。以合成的抗原肽免疫昆明系小鼠及兔，观察体液免疫反应。结果初步筛选出3种抗原指数较高的表位，即98～103aa（命名为B98），156～162aa（命名为B156），228～233aa（命名为B228）；合成的多重抗原肽经RP-HPLC层析后，纯度达到92.49％，MS鉴定其相对分子质量一致，误差小于0．1％。以合成的MAP疫苗免疫小鼠及兔后，可产生高滴度的特异性抗PCV2CAP抗体。结论PCV2CAP的B细胞新表位多重抗原肽有免疫原作用，将为PCV2CAP表位肽疫苗的研究奠定基础。
Porcine circovirus associated disease （PCVAD） encompasses a group of syndromes linked to infection with porcine circovirus type 2 （PCV2）. The only structural protein of viral capsid, Cap, has become the major target for developing PCV2 vaccines and serological diagnostic reagents. This study aimed to synthesize and identify the antigenic peptides of B cell epitope on the capsid protein （CAP） of porcine circovirus type 2 （PCV2）. The prediction of B cell epitopes of PCV2 CAP was conducted by DNAStar and BcePred software combining with amino acid average antigenic index method of WU Yu-zhang. A four-branch form of multiple antigen peptide （MAP） was synthesized which was linked a novel promiscuous T helper （Th） site for broad immunogenicity in multiple species. The four-branch MAP was synthesized by Fmoc method and analyzed by RP-HPLC and Mass- spectrum （MS）. Then, Outbred rabbit and Kunming mice were immunized with the synthesized MAP, respectively. And their humoral immune response was observed to evaluate the effect of the novel antigens in animals. The amino acids of N-terminal number 98-103 （named as B98）, 156-162 （named as B156） and 228-233 （named as B228） were predicted as the most potential epitopes and synthesized as immunogen. RP-HPLC revealed that the purity of synthetic MAP reached to 92.49%. The molecular weight was identified by MS, and the error was less than 0.1%. High titer of anti PCV2 CAP antibody was both obtained in rabbit and mice immunized with the synthesized MAP.The new epitopes B98, B156 and B228 confirm the MAP of B cell epitope on PCV2 CAP has immunogen effect, which will lay a foundation for the study of antigenic peptide vaccine of PCV2 CAP.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2014年第5期397-401,共5页
Immunological Journal
基金
重庆市科技攻关计划项目(CSTC
2011AB1078
2012GG-YYJS80014)
重庆市基础与前沿研究计划项目(cstc2013jcyjA10068)
重庆市教委科学技术研究项目(KJ130802)
