摘要
目的 :建立分泌抗HIV 1p2 4单克隆抗体 (mAb)的杂交瘤细胞株 ,并对其特性进行初步鉴定。方法 :以纯化的基因工程制备的p2 4抗原免疫BALB/c小鼠 ,取免疫小鼠的脾细胞与Sp2 /0骨髓瘤细胞融合 ,经HAT、HT选择培养及有限稀释法进行克隆化后 ,用间接ELISA法及Dotblot对其进行筛选和特性鉴定。结果 :筛选到 2株可分泌抗HIV 1p2 4mAb的杂交瘤细胞 ,其腹水效价为 1×10 -5,亲和力为 1.7× 10 4～ 1.8×10 4mol/L ,mAb的Ig亚类均为IgG1。两株mAb与HBcAg、HCVRNA阳性血清及HIVgp4 1等均无交叉反应 ,只与HIV 1p2 4抗原阳性血清产生特异反应。结论 :成功地建立了 2株可分泌抗HIV 1p2 4mAb的杂交瘤细胞 ,为进一步研制HIV 1p2
AIM: To set up hybridoma cells secreting monoclonal antibodies (mAbs) against HIV-1 p24 antigen and characterize their properties. METHODS: BALB/c mice were immunized with purified p24 protein and then the immunized mice’s splenocytes were fused with Sp2/0 cells. Monoclonal hybridoma cell lines were obtained by limiting dilution, HAT and HT-selective culture. The specificity of mAbs was identified by Dot blot and indirect ELISA. RESULTS: We had established two hybridoma cell lines secreting mAbs against HIV-1 p24 antigen. The titers of two mAbs in ascitic fluid were 1×10^(-5) and 1.7×10~4-1.8×10~4 mol/L, respectively. Both mAbs belonged to IgG1. Indirect ELISA detection demonstrated that both mAbs had no cross-reactivities with other viral antigens, such as HBcAg, HCV RNA positive sera and HIVgp41. CONCLUSION: Two hybridoma cell lines secreting mAbs against HIV-1 p24 antigen have been established successfully, which lays the foundation for detecting HIV-1 p24 antigen.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2004年第6期699-701,共3页
Chinese Journal of Cellular and Molecular Immunology
基金
军队"十五"项目基金资助 (No.0 1MB1 36
0 1Q1 37)
