非洲猪瘟病毒p54蛋白抗原表位预测及鉴定

B cell epitope prediction and identification on the p54 of African swine fever virus

为鉴定非洲猪瘟病毒(ASFV)p54蛋白的B细胞线性抗原表位,本研究利用生物信息学方法结合抗原指数对ASFV p54蛋白进行抗原表位的预测,人工合成预测的抗原表位多肽,利用ASFV阳性血清以及特异性单克隆抗体(MAb),采用间接ELISA和阻断ELISA方法验证所预测抗原表位多肽的免疫学特性。预测结果显示,可能的抗原表位区域有:aa23-aa29,aa36-aa45,aa72-aa94,aa114-aa120,aa124-aa130,aa137-aa150。间接ELISA试验表明aa23-aa29,aa36-aa45,aa72-aa94,aa114-aa120,aa137-aa150多肽的抗原性较好,其中aa36-aa45,aa72-aa94多肽可以分别与两株杂交瘤细胞分泌的MAb发生特异性反应,本研究为ASF多肽疫苗及免疫学临床诊断方法的建立奠定基础。

To identification of the B cell epitopes on the p54 of African swine fever virus,a series of short peptides were synthesized according to the amino acid sequence of p54 based on the hydrophilic and antigenic indexes for the epitope prediction by the software analysis.And their immunological characteristics were verified by indirect ELISA and blocking ELISA.The prediction results showed that antigen epitope regions were identified in aa23-aa29,aa36-aa45,aa72-aa94,aa114-aa120,aa124-aa130 and aa137-aa150.However,the five short peptides of aa23-aa29,aa36-aa45,aa72-aa94,aa114-aa120 and aa137-aa150 were verified to have antigenicity by indirect ELISA.And the short peptides of aa36-aa45 and aa72-aa94 were able to react with two monoclonal antibodies specifically.These results would facilitate the polypeptide vaccine development and the clinical immunology detection method establishment.