Fed-batch operation of an industrial cell culture process in shaken microwells

Abstract

Recently we have demonstrated batch suspension culture of mammalian cells in microwell plates. Here we describe a method for fed-batch culture of an industrially relevant GS-CHO (Glutamine Synthetase-Chinese Hamster Ovary) cell line in shaken 24-standard round well (24-SRW) plates. Use of a commercially available 'sandwich lid' and appropriate dilution of the bolus feeds counteracted liquid evaporation from the wells resulting in similar cell growth and antibody formation kinetics in both 24-SRW plates (800 mul) and shaken flasks (50 ml). Peak viable cell densities obtained were 8 +/- 0.5 x 10(6) and 9 +/- 1.3 x 10(6) ml(-1), respectively, while comparable final titres of a whole IgG of approximately 1.5 g l(-1) were recorded. Use of microwells provides at least a 50-fold reduction in medium requirements compared to shake-flask and other culture devices currently used in early stage cell culture process development. The ability to run multiple wells in parallel and to automate culture operation also offers considerable enhancements in experimental throughput.

CHO cell line specific prediction and control of recombinant monoclonal antibody N‐glycosylation

“…Caution must be exerted however, as the expression of a putative cell surface galactosyltransferase (Penno et al, ) and the presence of glycosidases (Porwoll et al, ) in the culture medium released by dead cells may influence terminal galactosylation of cell surface proteins, affecting the extent of correlation between cell surface lectin binding and MAb β1,4‐galactose levels. When taken in to account, by exploiting the advantages of cell surface glycan screening in a live‐cell lectin microarray format (Li et al, ; Tao et al, ; Tateno et al, ), there is potential to couple the technology to microscale cell culture systems and high throughput product titre measurement (Amanullah et al, ; Legmann et al, ; Silk et al, ) to rapidly identify culture conditions that result in cell lines with favourable productivity and desirable glycan processing. Rapid screening of cell lines in this way permits a level of feedback control, identifying promising production cell line candidates with ideal glycan processing early in development which can then undergo further manipulation during process optimisation.…”

High‐throughput screening of antibody‐expressing CHO clones using an automated shaken deep‐well system

“…Fed‐batch cultures were performed by feeding cells every 2 days with an in‐house feed solution. For the 96DW fed‐batch cultures, the inoculation and feeding processes were performed using the Hamilton robot (Hamilton, Reno, NV, USA), with nutrient feed being diluted with water to counteract reductions in volume due to evaporation . Only one sample was removed for titer or cell number determination during the fed‐batch in the 96DW plates and one sample was removed at the end.…”

Design and validation of a parallelized micro‐photobioreactor enabling phototrophic bioprocess development at elevated throughput

“…Moreover, growth phenotypes, likely induced by ventilation or temperature effects within the incubation chamber (Pacheco et al, ) or uneven CO 2 mass transfer into the wells (Han et al, ), were observed. With regard to extended cultivation times, evaporation control needs to be evaluated carefully, as high liquid loss may distort cultures’ performance (Betts et al, ; Silk et al, ). Many of the systems presented use top side illumination of the MTP potentially impeding the use of appropriate sterile barriers ensuring monoseptic conditions and restricting the accessibility with respect to integration into laboratory robotic platforms.…”