猪伪狂犬病毒变异株单克隆抗体的制备与鉴定

PREPARATION AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES AGAINST PORCINE PSEUDORABIES VIRUS VARIANTS

为制备猪伪狂犬病毒(Pseudorabies virus,PRV)变异毒株的单抗,以纯化灭活的PRV JS-2012毒株免疫BALB/c雌性小鼠。经间接免疫荧光法(indirect immunofluorescence assay,IFA)筛选,获得2株稳定分泌PRV单抗的杂交瘤细胞:4D8、4F10。2株单抗经亚型鉴定均为Ig G1亚类,轻链为κ链,腹水IFA效价均达1:51 200;IFA与IHC结果显示,二者均能与PRV毒株发生特异性反应;Western blot结果表明,4D8与PRV反应条带大小约为60 k Da,而4F10不与PRV发生反应,推测其可能针对PRV的构象性抗原表位。该单抗的制备对PRV的抗原变异及诊断研究均有重要意义。

In order to generate the monoclonal antibodies（MAbs） against a variant of Porcine pseudorabies virus（PRV）, BALB/c mice were immunized with purified and inactivated PRV JS-2012 strain. Two hybridomas secreting PRV specific MAbs were obtained and designated as 4D8 and 4F10, respectively. Both MAbs belonged to IgG1 subgroup and had κ light chains. The ascetic fluids containing either 4D8 or 4F10 showed indirect immunofluorescence assay（IFA） titers at 1：51200. Both MAbs reacted specifically with PRV in IFA and immunohistochemical（IHC）. In Western blot, MAb 4D8 reacted with 60 k Da protein of PRV while MAb 4F10 showed no obvious reactions with PRV protein, suggesting that MAb 4F10 might react with a conformational epitope. The availability of PRV MAbs was important for further research of antigenic variation and diagnosis.