Banik G G, Heath C A
Thayer School of Engineering, Dartmouth College, Hanover, New Hampshire 03755, USA.
Biotechnol Bioeng. 1995 Nov 5;48(3):289-300. doi: 10.1002/bit.260480315.
Steady state metabolic parameters for hybridoma cell line H22 were determined over a wide range of cell densities and specific growth rates in a filtration based homogeneous perfusion reactor. Operating the reactor at perfusion rates of 0.75, 2.0, and 2.9 day(-1)(each at four different specific growth rates), viable cell densities as high as 2 x 10(7) cells/mL were obtained. For the cell line under investigation, the specific monoclonal antibody production rate was found to be a strong function of the viable cell density, increasing with increasing cell density. In contrast, most of the substrate consumption and product formation rates were strong functions of the specific growth rate. Substrate metabolism became more efficient at high cell densities and low specific growth rates. The Specific rates of metabolite formation and the apparent yields of lactate from glucose and ammonia from glutamine decreased at low specific growth rates and high cell densities. While the specific oxygen consumption rate was independent of the specific growth rate and cell density, ATP production was more oxidative at lower specific growth rate and higher cell density. These observed shifts are strong indications of the production potential of high-density perfusion culture.
在基于过滤的均相灌注反应器中，在广泛的细胞密度和比生长速率范围内，测定了杂交瘤细胞系H22的稳态代谢参数。以0.75、2.0和2.9天⁻¹的灌注速率操作反应器（每个灌注速率对应四种不同的比生长速率），获得了高达2×10⁷个细胞/mL的活细胞密度。对于所研究的细胞系，发现特异性单克隆抗体产生速率是活细胞密度的强函数，随细胞密度增加而增加。相比之下，大多数底物消耗和产物形成速率是比生长速率的强函数。在高细胞密度和低比生长速率下，底物代谢变得更高效。在低比生长速率和高细胞密度下，代谢产物形成的比速率以及葡萄糖生成乳酸和谷氨酰胺生成氨的表观产率降低。虽然比耗氧速率与比生长速率和细胞密度无关，但在较低比生长速率和较高细胞密度下，ATP生成更多地通过氧化途径。这些观察到的变化强烈表明了高密度灌注培养的生产潜力。
