« BIOPROCESSING “
High-throughput process development and data analysis provides companies with the ability to perform more experiments and gain a greater understanding of the production process earlier in product development.
involved in biopharmaceutical manufacturing process development. These include determining the optimal temperature, pH, and DO for cell culture processes or the best load and elute conditions for a chromatography column used to purify a biopharmaceutical. A variety of micro- and mini-reactor formats for cell culture and fermentation optimization, plate-based, and micro-column-based products for the development of purification processes and robotic liquid handling systems are available from multiple industry suppliers. Together with the ever-increasing computational power of today’s systems, these tools can result in the collection and analysis of very large data sets. High-throughput process development and data analysis provides companies with the ability to perform more experiments and gain a greater understanding of the production process earlier in product development. This emerging trend is aligned well with increasing regulatory expectations and with the principles of Quality by Design that are becoming more widely applied to biopharmaceutical development.
chromatography media and evaluate over 1000 different operating conditions in the development and optimization of an entire MAb purification process. This comprehensive study was completed in just 63 days by 1 scientist, demonstrating the efficiency of the high- throughput methodology.
For the initial capture step, Lacki et al35 studied the relationship
between residence time and dynamic binding capacity to optimize the binding conditions for the antibody to MabSelect SuRe Protein A media, which was then verified in several small-scale column chromatography runs. In addition, the efficiency of different elution buffers was investigated using microtiter plates where it was found that the salt concentration of the elution buffer had no effect on either the overall step yield or monomer content of the recovered MAb. However, the overall yield and monomer content of the recovered MAb were affected by changes in the pH of the elution buffer. Decreases in pH increased the step yield up to 100% but also resulted in an increase in aggregate content.35
Following optimization of the Protein A capture step, aggregate removal by Capto Adhere media was evaluated using PreDictor plates from GE Healthcare. In an initial set of experiments, it was determined that both monomer and aggregates had bound to this media, making it impossible to obtain the desired yield and purity of the final product by running the column in a flow through mode. Therefore, the effect of salt type, salt concentration, and buffer pH on the efficiency of selective elution of monomer bound to the media was investigated, with conditions developed that allowed the recovery of monomeric antibody containing >1% aggregate. When these conditions were tested in a small-scale column, purified MAb containing >0.5% aggregate was recovered with a step yield of 87%.35
Summary
Recent technical advances have enabled a greater application of high- throughput methods to the traditionally labor-intensive activities
References
1. Flownamics® Web site. Available at:
http://www.flownamics.com/products_segflow.php. Accessed February 6, 2015.
2.
International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use Expert Working Group (CH). Pharmaceutical Development Q8(R2) Step 4. Geneva: International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use; 2009 Aug. 28 p. Available at:
http://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/ Quality/Q8_R1/Step4/Q8_R2_Guideline.pdf.
3. Rameez S, Mostafa SS, Miller C, Shukla AA. High-throughput miniaturized bioreactors for cell culture process development: reproducibility, scalability, and control. Biotechnol Prog. 2014 May-Jun;30(3):718-727.
4. Chhatre S, et al. A microscale approach for predicting the performance of chromatography columns used to recover therapeutic polyclonal antibodies. J Chomatogr A. 2009 Nov;1216:7806-7815.
5. Long Q, et al. The development and application of high-throughput cultivation technology in bioprocess development. J Biotechnol. 2014 Dec;192 Pt B:323-338.
6. Legmann R1, Benoit B, Fedechko RW, et al. A strategy for clone selection under different production conditions. Biotechnol Prog. 2011 May-Jun;27(3):757-765.
7. Molecular Devices. Cell Screening and Bioproduction Web Site. Available at: http://www.
moleculardevices.com/systems/cell-screening-and-bioproduction/clonepix-2-system. Accessed February 6, 2015.
8. 9.
Sartorius Stedim Biotech. TAP Biosystems Key Applications Web Site. Available at:
http://www.tapbiosystems.com/tap/applications/cell_line_selection.htm. Accessed February 6, 2015.
iClick Media. Bioprocessors Web Site. Available at:
http://www.iclickmedia.com/ bioprocessors/
system.htm. Accessed February 6, 2015.
10. Sartorius Stedim Biotech. Tap Biosystems Product Web Site. Available at: http://www.
tapbiosystems.com/tap/cell_culture/ambr.htm. Accessed February 6, 2015.
11. Sartorius Stedim Biotech. Tap Biosystems Product Web Site. Available at: http://www.
tapbiosystems.com/tap/cell_culture/ambr_250.htm. Accessed February 6, 2015.
12. Eppendorf. Benchtop Cell Culture Web Site. Available at:
http://eshop.eppendorfna.com/ products/DASGIP_Parallel_Bioreactor_Cell_Culture_RD. Accessed February 6, 2015.
13. Fogale Biotech Applications Web Site. Available at:
http://www.fogalebiotech.com/PHP/ applications-bioreactor.php. Accessed February 6, 2015.
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