分泌抗马铃薯X病毒株系特异性单克隆抗体杂交瘤细胞株的建立及抗体理化性质测定

SOME PROPERTIES OF POTATO VIRUS X SPECIFIC MONOCLONAL ANTIBODIES AND THE HYBRIDOMA CELL LINES

用马铃薯X病毒(PVX)免疫的BALB/c小鼠的脾细胞与骨髓瘤细胞(SP2/0)融合,经3次克隆化后,筛选出3类分泌抗PVX株系特异性的单克隆抗体(McAb)杂交瘤细胞株,该细胞株经传代20代以上,并经液氮冻存一年以后仍表现稳定。3个杂交瘤细胞株染色体数目集中在92—102条之间,其免疫球蛋白亚类均为IgG3,用ELISA间接法测定细胞培养上清滴度为1∶320—1∶640,小鼠腹水抗体滴度为1∶102400—1∶204800,后者比兔抗血清滴度(1∶320)高300倍以上。McAb对抗原具有中和作用,中和滴度为1∶10~2。经反复冻融、硫铵沉淀和冻干后,抗体活性无明显变化。

Seven hybridoma cell lines secreting monoelonal antibodies (McAbs) against potato virus X (PVX) were established by fusing mouse myeloma cells (SP2/0) with splenocytes of BALB/c mice immunized with purified PVX.One of the seven hybridoma cell lines secreting McAbs reacted specifically to a common strain of PVX, two reacted to a mild strain and the remaining four reacted to all of the five strains tested. Three hybridoma cell lines, namely 1-1H5, 2-2A7, and 1-4C6 secrete IgG3 immunoglobulins. The McAb titers of ascitie fluids were about 300 times of that of rabbit antisera measured by indirect ELISA. The MeAbs had a 1:10~2 neutralization titer. The reactivities of McAbs were not affected significantly by freeze drying, repeatedly freezing-thawing, or precipitating with ammonium sulphate.