摘要
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.
出处
《生物工程学报》
CAS
CSCD
北大核心
1989年第2期108-112,共5页
Chinese Journal of Biotechnology
基金
国家科委生物工程开发中心资助