抗鸡传染性法氏囊病病毒VP3单克隆抗体制备及抗原表位的初步分析

Development of Monoclonal Antibodies against VP3 of IBDV and Identification of Antigenic Epitopes

用纯化的原核表达的IBDVVP3蛋白免疫6～8周龄的雌性BALB／c小鼠，经过3次免疫后，取其脾细胞与SP2／0骨髓瘤细胞进行融合，经过3次有限稀释，获得分泌抗IBDVVP3抗体的4株（HRB-7B、HAR-7C、HRB-3F、HRB-10E）杂交瘤细胞系。结果表明，4株杂交瘤细胞分泌的抗体均能够特异地与IBDVVP3蛋白反应，细胞上清EuSA效价均在5．0×10^2以上，腹水EuSA效价为6．4×10^6以上；相加ELISA及肽扫描结果表明，4株中仅HRB-7C和HRB-10E2株单抗针对的抗原表位相近，并利用肽扫描的方法初步定位4株单克隆抗体的抗原表位。本研究对进一步分析IBDV的结构与功能以及建立以表位为基础的抗原抗体诊断方法具有重要的意义。

Female, 6-8 weeks old BALB/c mice were immunized with the purified recombinant fusion VP3 of IBDV. After three immunizations, the spleen cells from immunized BALB/c mice were fused with mouse myeloma cells SP2/0. By 3 cycles of limited dilutions, we obtained four strains of hybridoma cells, named HRB-TB, HAR-TC, HRB-3F and HRB-10E. Isomerism and stability analysis indicated that those monoclonal antibodies had specific reaction with the VP3 of IB- DV. The titer of four antibodies were up to 5.0× 10^2 in cell culture supernatants and 6.4× 10^6 in ascetic fluids. Plusing ELISA showed that in four cell strains, HRB-TC and HRB-10E have the similar antigen epitopes. Moreover, the study identified the antigenic epitopes by peptide scanning, further structural and function analysis of IBDV. This work could provide the basis for the development of immunity-based prophylactic, therapeutic and diagnostic techniques for the control of IBD and further structural and function analysis of IBDV.