A型流感病毒M2e单克隆抗体的研制及鉴定

Preparation and Characterization of Monoclonal Antibodies Recognizing SLLTEVETPIRNEWGSRSNDSSD Epitope of Influenza A Virus M2 Protein

目的制备A型流感M2e(2-24)单克隆抗体,并进行初步研究。方法以M2e-OVA免疫BALB/c小鼠,取免疫后的小鼠脾细胞和SP2/0细胞融合,经有限稀释法克隆化及筛选,获得抗M2e(2-24)的杂交瘤细胞株,制备腹水,并对其进行纯化,分析和鉴定。结果获得3株稳定分泌M2e单克隆抗体的杂交瘤细胞株L1,L2,L3,其免疫球蛋白亚类均为IgG1,腹水效价为105以上。结论成功制备了抗M2e(2-24)的特异性单克隆抗体,为研制具有交叉保护作用的流感疫苗奠定了基础。

Objective To prepare monoclonal antibodies （McAbs） against M2e （2 - 24）, and to study its character. Methods BALB/c mice were immunized with M2e - OVA. Spleen cells of the mice were infused with SP2/0 myeloma cells. Three strains of high affinity hybridomas and McAbs against M2e were screened by ELISA and cloned by limiting dilu- tion. The ascites was prepared, purified, and analyzed. Results Three strains of hybridomas were obtained, which were able to secrete McAbs stably. The immunoglobulin subclass was IgG~. Their ascites titers were all above 105. Conclusions The high affinity McAbs against M2e are produced by the ten strains of hybridomas, which lay a foundation for detection of influnza A by ELISA.