Mochizuki K, Sato S, Kato M, Hashizume S
Morinaga Institute of Biological Science, Yokohama, Japan.
Cytotechnology. 1993;13(3):161-73. doi: 10.1007/BF00749812.
It was found that the production of human monoclonal antibodies (MoAbs) by human-human hybridomas can be significantly enhanced by replacing glucose with fructose in the dish culture medium. Optimization of initial concentrations of fructose and glutamine, another influencing factor for MoAb production, enabled an enhanced production of human MoAb 2.1 times higher than that obtained using the conventional culture media employing glucose. It was shown by kinetic analysis that enhanced MoAb production at the optimum fructose concentration can be attributed to the retention of high specific antibody production rates and diminished time lag during the course of culture. These dish culture results with fructose-containing medium were successfully applied to the continuous perfusion culture with a slight modification, where 2.9- and 1.9-fold enhancements in specific antibody production rate and MoAb concentration, respectively, were attained as compared with the conventional glucose-containing medium. An inverse relationship was observed between the secreted concentrations of lactic acid and MoAb when the hybridoma was cultured in the media containing varying concentrations of fructose, i.e., the lower the lactic acid concentration, the higher the MoAb production and vice versa, suggesting that fructose at appropriate concentrations in the medium can serve as an alternative sugar for the efficient production of human MoAbs, with reduced pH shifts, for the serum-free culture of human-human hybridomas.
研究发现，在培养皿培养基中用果糖替代葡萄糖可显著提高人-人杂交瘤产生人单克隆抗体（MoAbs）的产量。优化果糖和谷氨酰胺的初始浓度（另一个影响MoAb产生的因素）后，人MoAb的产量比使用含葡萄糖的传统培养基提高了2.1倍。动力学分析表明，在最佳果糖浓度下MoAb产量的提高可归因于高特异性抗体产生率的保持以及培养过程中时间滞后的减少。这些含果糖培养基的培养皿培养结果经轻微修改后成功应用于连续灌注培养，与传统含葡萄糖培养基相比，特异性抗体产生率和MoAb浓度分别提高了2.9倍和1.9倍。当杂交瘤在含有不同浓度果糖的培养基中培养时，观察到乳酸分泌浓度与MoAb之间呈反比关系，即乳酸浓度越低，MoAb产量越高，反之亦然，这表明培养基中适当浓度的果糖可作为替代糖，用于高效生产人MoAbs，减少pH值变化，用于人-人杂交瘤的无血清培养。
