SPA单克隆抗体的制备与纯化

Preparation and Purification of the SPA-Specific Monocloal Antibody

以SPA作基础免疫BALB/C小鼠后,融合前三天直接脾内免疫,取脾细胞与SP_2/O瘤细胞常规融合,融合率100％;以快速EIISA检测抗体,阳性率60％。5次有限稀释法克隆化后获得7株杂交瘤细胞,作中和抑制试验,证实为SPA特异性抗体。经鉴定属IgG2a及IgG2b亚类。电镜观察细胞融合,发现细胞内含逆转录病毒。经MAPS—100法纯化单抗腹水,获得了高纯度McAb。

The Balbc mice were primarily immunized with SPA, and then were boosted intraspleenly with the same antigen 3 days prior to cell fusion. The hybridoma cells were made by fusing the immunized mouse spleen cells with SP2/0 myeloma cells inuconventional fusingfashion. The fusing rate was 100%. The antibody secretimg cells were screened by a guick ELISA method. The percentage of positive clones was 60%.. Through 5 times of limiting dilution cloning. The 7 strain of hybridoma cells were established. The antibodies produced by these cells were confirmed to be SPA-specific by the specific neutralization assay, purtheremore, the antibodies were identified to belong to two s-uclasses of immunoglobulins: IgG2a and IgGsb respectively. The retro-virus particles were detected in the cytoplasm of the hybridoma cells by electronic microscope examination. Finally, the SPA-specific antipodes were highly purified from the ascites fluid by a MAPS-100 system (BIO-RAD).