Automated Concentration and Diafiltration of Multiple siRNA Samples - A team of scientists from GE Healthcare collaborated with Merck scientists on a project to transfer a conventional manual concentr

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Automated Concentration and Diafiltration of Multiple siRNA Samples
A team of scientists from GE Healthcare collaborated with Merck scientists on a project to transfer a conventional manual concentration/diafiltration process for siRNA production.


BioPharm International
Volume 26, Issue 2, pp. 22-26

RESULTS


Figure 5: Example siRNA DF screening result. Left axes are ConcFactor in blue and DF X Factor in red. Right axes are LMH in green and Feed Pressure in blue. DF=diafiltration factor, LMH=permeate flux (L/m2/hour).
Up to five samples/day/system were processed with start up time of less than one hour. The chromatogram in Figure 5 illustrates a portion of the data observed for the processing of a siRNA sample. Event marking indicates the various segments of the method run. The curve data track and record all information in an electronic record. Curve data not depicted are automatically included in the result file and can be presented at a later date if desired.

CONCLUSION


Table II: Advantages of automated versus manual system setup. FTE is full-time employee.
Automating the ultrafiltration step of siRNA production increased the overall productivity of the group by 40% and reduced the necessary manpower required for the project from two full-time employees (FTEs) to one. The 40% improvement was a result of being able to process 10 samples/day on two automated systems compared with the previous approach of processing two samples/day on three manual systems (see Table II).

These improvements allowed for overall cost savings while increasing the number of samples available for research. By utilizing the instructions available in the UNICORN software for the ÄKTAcrossflow platform, a safety measure was incorporated into the method to prevent loading sample onto a fouled filter, reducing the risk of product lost and failed experiments. The ÄKTAcrossflow system has proven to be suitable for automated multiple sample processing using standard software and hardware with minimal operator interaction, demonstrating an efficient means to improved process robustness.

Rebecca Arvary is a research chemist at Merck Research Labs, Merck & Co, RY818-B221, PO Box 2000, Rahway, NJ 07065 USA. *Deborah R. Cohen is a senior scientist of Fast Trak, Steven Vaughan is a field applications consultant, and Catherine Blake is a product specialist-filtration, all at GE Healthcare Life Sciences, 800 Centennial Avenue, Piscataway, NJ 08854 USA.

*To whom all correspondance should be addressed
.

PEER-REVIEWED

Article submitted: Aug. 22, 2012. Article accepted: Oct. 12, 2012.

REFERENCES

1. T. Tokatlian and T. Segura, WIREs Nanomed. Nanobiotechnol. 2 (3), 305–315 (2010).

2. R.S. Pellish et al., Pharmacol. Ther. 27 (9), 715–723 (2008).

3. O. Pontes and C.S. Pikaard, Curr. Opin. Genet. Dev. 18 (2), 197–203 (2008).

4. M. Schlee et al., Mol. Ther. 14 (4), 463–470 (2006)

5. M. Ichihara et al., Nucl. Acids Res. 35 (18), e123 (2007).


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