We would like to thank Drs. Chris Curtin and Simon Conn, Flinders University for technical assistance with the HPLC analysis. “
“The demand for antibodies and other glycoproteins has increased rapidly due to their importance as therapeutic agents. Production of these biologics
is in large part reliant on mammalian cell culture systems. To meet the demand, various strategies are employed to increase production by enhancing cell performance, specifically improving bioproduction titers. Some of the most effective approaches to improve the cell culture process include media optimization and batch and fed batch supplementation. Development of these strategies involves intensive screening of components, mixtures and various feed schemes. Due to the difficulty of examining a large number of components at different concentrations or screening a large PLX4032 number of potential supplements, these types of studies cannot be efficiently performed using conventional bioreactor, shake flask, or spin tube cultures. Use of small scale cell culture systems, like multi-well plates can enable the exploration of a wider range of bioprocess operating conditions in a more efficient manner. In general, multi-well plate cell cultures are performed in static or shaking formats in shallow well (SW) or deep well (DW) plates in a small culture volume for elongated time periods. Shaking plates
are more suitable for suspension cell culture; however, the loss of media due to evaporation in outer wells Thalidomide (edge effects) poses a significant problem [1]. Edge effects result in well-to-well and plate-to-plate variability in cultures. Erastin Some laboratories have overcome the edge effects and evaporation issues by not using the outer wells of multi-well plates for cell-based assays. Omission of outer wells decreases analysis throughput by 38% and 66% in 96 well and 24 well plates, respectively. Other approaches to prevent evaporation
include the use of self-adhesive plate seals, which are designed to maintain uniform gas exchange while keeping cultures sterile. A study of various commercially available adhesive plate seals showed multiple disadvantages and categorized them into two groups: (1) plate seals in which volume preservation is relatively low, but oxygen transfer is comparable to that of unsealed plates, and (2) plate seals in which volume preservation is high, but oxygen exchange is slower [2]. Therefore, adhesive plate seals may not adequately fulfill the requirements for cell culture screening in SW and DW multi-well plates. In addition to evaporation and oxygen transfer issues, culture volumes in most of the 24SW, 96SW, and 96DW plates ranges from 0.7 mL to 1 mL. These lower volumes are usually insufficient for multiple sampling and fed batch processes. Moreover, for suspension cultures, plates are being kept on shaking platform to keep cells in suspension. It is often challenging to keep plates secure on a regular platform while shaking.
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