Gaertner J G, Dhurjati P
Department of Chemical Engineering, University of Delaware, Newark 19716.
Biotechnol Prog. 1993 May-Jun;9(3):298-308. doi: 10.1021/bp00021a009.
A comprehensive approach was taken toward the quantitative study of hybridoma growth and antibody production. A fractional factorial experimental method was used to identify important variables and variable interactions affecting hybridoma behavior. The variables studied include temperature, pH, dissolved oxygen, glucose, glutamine, base medium, serum, lactate, and ammonium. The growth rate was strongly affected by the levels of dissolved oxygen, pH, temperature, and base medium. Interactions between temperature, pH, and dissolved oxygen were important. The optimal pH for growth depends upon the temperature and dissolved oxygen concentration. In general, growth was fastest at low dissolved oxygen levels. The growth rate was very sensitive to low concentrations of base medium, but was relatively insensitive to the serum concentration at levels above 2.5%. Antibody production was stimulated by high concentrations of base medium and serum and inhibited by ammonium and lactate. Antibody production increased linearly with serum concentration. In general, conditions that favored a high growth rate also favored a high specific rate of antibody production. The functional dependencies of antibody production on base medium and ammonium were similar to those for cell growth; however, antibody production and cell growth exhibited different dependencies on serum. Mathematical descriptions of cell growth and antibody production were developed. These experimental results have significant implications for the optimization of hybridoma growth in bioreactors.
我们采用了一种综合方法来对杂交瘤生长和抗体产生进行定量研究。使用分数析因实验方法来确定影响杂交瘤行为的重要变量及其相互作用。所研究的变量包括温度、pH值、溶解氧、葡萄糖、谷氨酰胺、基础培养基、血清、乳酸和铵。生长速率受到溶解氧水平、pH值、温度和基础培养基的显著影响。温度、pH值和溶解氧之间的相互作用很重要。生长的最佳pH值取决于温度和溶解氧浓度。一般来说，在低溶解氧水平下生长最快。生长速率对低浓度的基础培养基非常敏感，但对血清浓度在2.5%以上时相对不敏感。高浓度的基础培养基和血清会刺激抗体产生，而铵和乳酸则会抑制抗体产生。抗体产生随血清浓度呈线性增加。一般来说，有利于高生长速率的条件也有利于高特异性抗体产生速率。抗体产生对基础培养基和铵的功能依赖性与细胞生长相似；然而，抗体产生和细胞生长对血清表现出不同的依赖性。我们建立了细胞生长和抗体产生的数学描述。这些实验结果对生物反应器中杂交瘤生长的优化具有重要意义。
