抗肠道病毒EV71型外壳蛋白VP1单克隆抗体的制备和鉴定

目的 制备具有中和活性的抗肠道病毒EV71型外壳蛋白VP1的单克隆抗体.方法 人工合成SP55和SP70(分别包含VP1的第163-177,208-222位氨基酸)两段VP1的多肽,分别免疫BALB/c小鼠,常规杂交瘤技术进行细胞融合,用间接酶联免疫吸附试验(ELISA)筛选阳性杂交瘤细胞并测定效价.用分泌的单抗和EV71病毒在RD细胞上进行中和试验以检验其中和活性.结果 得到2株能稳定分泌抗肠道病毒EV71型VP1蛋白单克隆抗体的杂交瘤细胞株,2株单抗的中和效价分别为1:8和1:16.结论 成功制备出2株具有中和活性的抗肠道病毒EV71型VP1蛋白单克隆抗体,为其下一步应用打下基础.

Preparation and identification of the monoclonal antibodies against VP1 capsid protein of Enterovius 71

Objective To prepare monoclonal antibodies (McAbs) against VP1 capsid protein of Enterovims 71. Methods Two peptides, SP55 and SPT0, containing amino acid 163-177 and 208-222 of VP1, were synthesized respectively. Immunized BALB/c mice with the synthetic peptides to estabhsh the hybridoma cell strains secreting specific McAb to VP1. After the specific McAbs were prepared, identified and analyzed the titer by indirect ELISA assay. The positive clones were selected and their neutralization titer were determined by neutralization test. Results Two high titered anti-VP1 antibodies secreted by the hybfidoma cells showed good neutralization reaction with enterovirus 71 on RD cells, and the neutralization titer were 1:8 and 1 : 16 respectively. Conclusion Two high titered anti-VP1 antibodies, with good neutralization activity, secreted by the hybridoma cells, which lays the foundation for further study.