Use of disposable reactors to generate inoculum cultures for E. coli production fermentations

Abstract

Disposable technology is being used more each year in the biotechnology industry. Disposable bioreactors allow one to avoid expenses associated with cleaning, assembly and operations, as well as equipment validation. The WAVE bioreactor is well established for Chinese Hamster Ovary (CHO) production, however, it has not yet been thoroughly tested for E. coli production because of the high oxygen demand and temperature maintenance requirements of that platform. The objective of this study is to establish a robust process to generate inoculum for E. coli production fermentations in a WAVE bioreactor. We opted not to evaluate the WAVE system for production cultures because of the high cell densities required in our current E. coli production processes. Instead, the WAVE bioreactor 20/50 system was evaluated at laboratory scale (10-L) to generate inoculum with target optical densities (OD(550)) of 15 within 7-9 h (pre-established target for stainless steel fermentors). The maximum settings for rock rate (40 rpm) and angle (10.5) were used to maximize mass transfer. The gas feed was also supplemented with additional oxygen to meet the high respiratory demand of the culture. The results showed that the growth profiles for the inoculum cultures were similar to those obtained from conventional stainless steel fermentors. These inoculum cultures were subsequently inoculated into 10-L working volume stainless steel fermentors to evaluate the inocula performance of two different production systems during recombinant protein production. The results of these production cultures using WAVE inocula showed that the growth and recombinant protein production was comparable to the control data set. Furthermore, an economic analysis showed that the WAVE system would require less capital investment for installation and operating expenses would be less than traditional stainless steel systems.

Fluid dynamic characterization of a laboratory scale rocked bag bioreactor

“…The scale limitation of commercially available rocked bags (currently the maximum working volume is 500 L, Wave 1000/500 by GE Healthcare) means they are used mainly used as seed vessels and for clinical trial material production. Several studies have been carried out in rocked bags using a variety of organisms such as S. cerevisiae , Baculovirus , plant cells, and E. coli . Rocked bag systems have also been operated in perfusion mode and for microcarriers suspension in stem cell expansion studies .…”

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

“…The operating cost is high at large scale due to frequent purchasing of new bags. Single-use cultivation systems are also utilized to prepare inoculum for large-scale cultivations (Mahajan et al, 2010).…”