High‐End pH‐controlled delivery of glucose effectively suppresses lactate accumulation in CHO Fed‐batch cultures

Abstract

A simple method for control of lactate accumulation in suspension cultures of Chinese hamster ovary (CHO) cells based on the culture's pH was developed. When glucose levels in culture reach a low level (generally below 1 mM) cells begin to take up lactic acid from the culture medium resulting in a rise in pH. A nutrient feeding method has been optimized which delivers a concentrated glucose solution triggered by rising pH. We have shown that this high-end pH-controlled delivery of glucose can dramatically reduce or eliminate the accumulation of lactate during the growth phase of a fed-batch CHO cell culture at both bench scale and large scale (2,500 L). This method has proven applicable to the majority of CHO cell lines producing monoclonal antibodies and other therapeutic proteins. Using this technology to enhance a 12-day fed-batch process that already incorporated very high initial cell densities and highly concentrated medium and feeds resulted in an approximate doubling of the final titers for eight cell lines. The increase in titer was due to additional cell growth and higher cell specific productivity.

Generic Raman‐based calibration models enabling real‐time monitoring of cell culture bioreactors

“…10,11 Regulators continue to require increasingly detailed and complex measurement of processes. Underfeeding will deprive the cells of an important nutrient and their main source of energy, while overfeeding leads to conversion of excess nutrients to lactate, which is an inhibitor of cell productivity.…”

Proteomic Analysis of Bioreactor Cultures of an Antibody Expressing CHOGS Cell Line that Promotes High Productivity

“…The two bioreactors that had the highest viability [R10H021 and R10H022] were initiated with lower amounts of the growth enhancing supplement F8 to slow down the initial rate of metabolism. This strategy of reduced feeding has been successfully tested previously [9,10]. In the current study, the reduced feeding is subsequently augmented by an increase in BRX feeding, such that high cell density is achieved.…”

“…To meet this demand, a variety of strategies have been employed to develop efficient biopharmaceutical manufacturing processes. These strategies include: 1) maximizing the productivity of the manufacturing cell line by the judicious use of expression systems for optimal transcription and translation of the therapeutic protein [2]; 2) engineering the transfection host cell line for efficient post-translational modification and secretion [3]; and 3) improving the cell culture process including media optimization [4][5][6][7][8], developing advanced feeding strategies [9,10] which in turn increases the culture density. Additionally, robust and highly productive host cell lines can be deployed by increasing the efficiency of gene expression that regulates proliferation [11] survival and longevity [12][13][14].…”

Mammalian Expression System and Improvisation for High Production

“…By using the high end pHcontrolled feeding strategy for the lactate effect, the mAb and other recombinant protein production titers has been doubled in almost all CHO cell lines from bench to large scale production [20].The fed batch process with the highly defined chemical media reportedly increased the production rate from 44 p/c/d to 54 p/c/d using plant hydrolysate and the maximum titer of 10 -13 g/L of mAb and Fc-fusion protein [19]. Raman spectroscopy is used to analyze and maintain the consistency of the media components for large scale production [21].…”