Cell Culture Clone Selection

Abstract

 

Effects of temperature-shift on cell line selection in recombinant Chinese hamster ovary (CHO) fed-batch cultures

Andrew Ray*¹, An Zhang², Chris Kwiatkowski², Yao-Ming Huang²

1. Chemical Engineering, North Carolina State University, Raleigh, NC; 2. Cell Culture Development, Biogen, RTP, NC       

In the biopharmaceutical industry, a temperature-shift process is widely used to improve cellular performance and product yields of recombinant proteins in mammalian cell culture, especially recombinant Chinese hamster ovary (CHO) cells. However, much less work has been published regarding the impact of the temperature-shift to sub-physiological temperature on the cell line selection, clone ranking. In the described work, we compared the relative clone ranking of a traditional single temperature fed-batch process to a temperature-shift fed-batch process. In that the cellular response to such a temperature shift is both cell- and product-specific, we first identified critical process parameters and optimal processing ranges for a temperature-shift process using Biogen’s cell culture platform. Then we successfully applied the temperature-shift fed-batch process to cell line selections of two different mAbs, using the high-throughput ambr15 system. The results indicated that the relative clone ranking among the selected subclones of the temperature-shift process were comparable to the single temperature process. Additionally, relative to the single temperature process the temperature-shift process increased harvest viability, reduced total feed volume, maintained or improved titer and improved product quality attributes. In a proceeding study, the previous results were reaffirmed in bench top bioreactors.