Xiao X W, Guo J, Cai S H, Liu J, Feng L X, Hsu H T
Biotechnology Research Center, Chinese Academy of Agricultural Sciences, Beijing.
Chin J Biotechnol. 1989;5(2):89-95.
Splenocytes from BALB/c mouse immunized with potato virus X (PVX) and myeloma cells (SP/2) were fused. After cloning and three screening cycles, three hybridoma cell lines secreting strain-specific monoclonal antibodies (MCAs) against PVX were obtained. These cell lines were stable for 20 generations and after storage in frozen form (in liquid nitrogen) for one year. The chromosome numbers of the three hybridoma cell lines clustered in the 92-102 range. The MCAs all belonged to the IgG3 immunoglobulin subclass. The medium supernatant antibody titer detected by indirect ELISA was 1:320-1:640. The mouse ascites antibody titer was 1:102,400-1:204,800, which was more than 300 times the rabbit antiserum titer (1:320). The MCAs had a neutralizing effect on the antigen, with neutralization titer of 1:10(2). There were no apparent changes in antibody activity after repeated freezing/thawing cycles, ammonium sulfate precipitation, or freeze-drying.
将用马铃薯X病毒(PVX)免疫的BALB/c小鼠的脾细胞与骨髓瘤细胞(SP/2)进行融合。经过克隆和三个筛选周期后,获得了三株分泌针对PVX的株系特异性单克隆抗体(MCA)的杂交瘤细胞系。这些细胞系在传代20次以及在冷冻状态(液氮中)保存一年后仍保持稳定。这三株杂交瘤细胞系的染色体数聚集在92 - 102范围内。这些MCA均属于IgG3免疫球蛋白亚类。通过间接ELISA检测,培养基上清液抗体效价为1:320 - 1:640。小鼠腹水抗体效价为1:102,400 - 1:204,800,是兔抗血清效价(1:320)的300多倍。这些MCA对抗原有中和作用,中和效价为1:10²。在反复冻融循环、硫酸铵沉淀或冻干后,抗体活性没有明显变化。