Novokhatskiĭ A S, Malakhova I V, Mikheeva T G
Vopr Virusol. 1985 Sep-Oct;30(5):602-8.
Hybridomas producing monoclonal antibodies to tick-borne encephalitis, Venezuelan equine encephalomyelitis, and vaccinia viruses were cloned and recloned by the end-point dilution method in 96-well Linbro plates on a cell feeder layer. With decreasing of the cell seed dose the average number of clones per well decreased and the effectiveness of clone production increased. When an average of one cell per well was introduced, the portion of vells with single clones for hybridomas of various origins ranged from 5.4% to 37.5%. As a rule, the first cloning resulted in a significant rise in titres of monospecific immunoglobulins secreted by hybridomas and improved growth characteristics of the hybrid cell population. Repeated cloning required in most cases for stability of the hybrid line was usually not accompanied by changes in the productivity of the cell cultures.
通过终点稀释法,在细胞饲养层的96孔林布罗板中对产生抗蜱传脑炎病毒、委内瑞拉马脑炎病毒和痘苗病毒单克隆抗体的杂交瘤进行克隆和再克隆。随着细胞接种剂量的降低,每孔克隆的平均数量减少,克隆产生的效率提高。当每孔平均接种一个细胞时,不同来源杂交瘤的单克隆孔比例在5.4%至37.5%之间。通常,第一次克隆会导致杂交瘤分泌的单特异性免疫球蛋白滴度显著提高,并改善杂交细胞群体的生长特性。大多数情况下,为了使杂交瘤系稳定而需要进行的重复克隆通常不会伴随着细胞培养物生产力的变化。