Institute of Applied Microbiology, University of Agriculture, Nussdorfer Lände 11, A-1190, Vienna, Austria.
Cytotechnology. 1995 Jan;17(2):83-9. doi: 10.1007/BF00749395.
Human-mouse heterohybridomas producing human monoclonal antibodies show slower growth rates and lower peak cell densities than murine hybridomas. In order to improve the growth properties we transfected a heterohybridoma cell line with expression plasmids containing the oncogenes v-src, c-Ha-ras and SV40largeT. The plasmids were transferred by electroporation. Growth promoting activities of the plasmids were proven in NIH3T3 cells whereby a doubling of the maximum cell densities of this cell type was observed. The oncogene products were analyzed by means of northern blotting and immunofluorescence. After transfection of c-src and c-ras, a heterohybridoma cell line was derived which showed improved growth characteristics compared to the original cell line. Although specific antibody production was lower, antibody concentrations which accumulated in batch culture were higher due to increased maximum cell densities.
产生人单克隆抗体的人鼠异源杂交瘤的生长速度比鼠源杂交瘤慢，峰值细胞密度也较低。为了改善生长特性，我们通过电穿孔将表达质粒转染入异源杂交瘤细胞系，这些质粒包含癌基因 v-src、c-Ha-ras 和 SV40largeT。在 NIH3T3 细胞中证明了质粒的促生长活性，观察到该细胞类型的最大细胞密度增加了一倍。通过 northern blot 和免疫荧光分析癌基因产物。转染 c-src 和 c-ras 后，获得了一株异源杂交瘤细胞系，与原始细胞系相比，该细胞系的生长特性得到了改善。尽管特异性抗体产量较低，但由于最大细胞密度增加，批量培养中积累的抗体浓度更高。
