抗猪瘟兔化弱毒单克隆抗体杂交瘤细胞株的建立

用猪瘟兔化弱毒（ＳＦＶ—Ｃ）牛睾丸细胞培养上清液，经ＰＥＧ—２００００沉淀和蔗糖密度梯度离心提纯制备ＳＦＶ—Ｃ抗原，腹腔免疫Ｂａｌｂ／ｃ小鼠，取抗体阳性的免疫小鼠脾细胞，在５０％（Ｗ／Ｖ）ＰＥＧ—４０００作用下，与ＳＰ２／０－Ａｇ１４小鼠骨髓瘤细胞进行融合，产生杂交瘤细胞。用间接ＥＬＩＳＡ法筛选阳性杂交瘤细胞株。通过有限稀释进行３次克隆，获得了２株分泌抗ＳＦＶ—Ｃ的单克隆抗体杂交瘤细胞株，并对其染色体形态和数目进行分析鉴定。

ESTABLISHMENT OF HYBRIDOMA CELL LINES SECRETING MONOCLONAL ANTIBODIES AGAINST LAPINIZED CHINESE STRAIN OF SWINE FEVER VIRUS

Lapinized chinese strain of swine fever virus (SFV-C) from the supernatant of bovine testis cell culture was concentrated by precipitation with polyethylene glycol(M.W.20000) and purified by sucrose density gradient centrifugation. Balb/c mice were inoculated three times intraperitoneally with SFV-C purified. Hybrid cells were produced by fusion of spleen cells of Balb/c mouse with positive serum and mouse myeloma cells SP2/0-Ag14 using 50% (W/V) PEG (M.W.4000). Positive hybridoma cells were selected by screening their supernatants with an indirect enzyme-linked immunosorbent assay.Two hybridoma cell lines secreting monoclonal antibodies against SFV-C were estalished after 3 clonings by limiting dilution. The morphologieal features as well as the numbers of the hybridomas nucleic chromosomes were identified.