A Purification Platform for the Production of MAbs from Fermenters with Titers of 5 g/L and Beyond - New techniques can overcome bottlenecks in existing facilities. - BioPharm International

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A Purification Platform for the Production of MAbs from Fermenters with Titers of 5 g/L and Beyond
New techniques can overcome bottlenecks in existing facilities.


BioPharm International Supplements


Cation Exchange Membrane Adsorbers

After the limitation of the Protein A wash buffer is removed, time again becomes a limiting factor. The longest processing time at high titers in this example will become the final polishing step to remove aggregates (performed with a high flow rate cation exchange resin). A membrane adsorber was tested to overcome this issue. Membrane adsorbers have previously been used in MAb manufacture for the removal of impurities and viruses, so the precedent exists to use this type of technology in established manufacturing processes.9 Because the binding capacity of adsorbers is less than that of chromatography resins, it was decided to develop the process in flow-through mode. This had the added advantage of reducing buffer volumes.


Figure 2. The variation in process performance with changing sodium chloride concentration in the feed stream
Capacities were high compared to chromatography, and flow rates increased by more than an order of magnitude, so the processing time of the unit operation was greatly reduced. Step yields and aggregate clearance were comparable, if not better than those seen with the high flow rate resin (Figure 2). The use of this technique combined with HP-TFF enabled the purification of fermentation batches of 9.1 g/L titer with no bottlenecks.

Conclusions


Figure 3. A graphical representation of how implementing technologies and techniques have and can increase the capabilities of downstream processes to purify fermenters with titers of over 9 g/L.
Improvements in downstream process design have enabled significant increases in plant throughput to be achieved. Limitations can still exist, but the implementation of existing technologies can improve the capability of the purification process still further. It is therefore possible to purify the output of even the largest scale fermenter to beyond 5 g/L, even approaching 10 g/L (Figure 3), by the implementation of techniques without the need for radical process re-design or expensive retrofitting of existing manufacturing facilities.

JIM DAVIES is the principal group leader in purification development at Lonza, Berkshire, UK (headquartered in Basel, Switzerland), +44 0 1753 716634,

References

1. Buckland, BC, Lilly MD. Fermentation: an overview. In: Stephanopoulos X, editor. Biotechnology, 2nd edition, volume 3. New York: Wiley-VCH; 1993.

2. Jagschies, G. Antibody purification from high titre cell lines. Bioproduction. Dublin; 2006 Oct 24–6.

3. Hagel L, Jagschies G, Sofer G. Handbook of process chromatography: Development, manufacturing, validation, and economics, 2nd edition. London: Elsevier; 2008.

4. De los Reyes G, Mir L, inventors; SPF Innovations, assignee. Method and apparatus for the filtration of biological solutions. United States patent US 7384549. 2008 Jun 10.

5. Van Reis R, Gadam S, Frautschy LN, Orlando S, Goodrich EM, Saksena S, Kuriyel R, Simpson CM, Pearl S. Zydney AL. High performance tangential flow filtration. Biotechnol Bioeng. 1997;56:71–82.

6. Ghosh, R. Protein Bioseparation Using Ultrafiltration; Theory, Applications and New Developments. London: Imperial College Press; 2003.

7. Wan Y, Ghosh R, Hale G, Cui Z. Fractionation of bovine serum albumin and monoclonal antibody Alemtuzumab using carrier phase ultrafiltration. Biotechnol Bioeng. 2005;90:303–15.

8. Christy C, Adams G, Kuriyel R, Bolton G, Seilly A. High-performance tangential flow filtration: a highly selective membrane separation process. Desalination. 2002;144:133–6.

9. Zhou JX, Tressel T. Basic concepts in Q membrane chromatography for large-scale antibody production. Biotechnol Progr. 2006;22:341–349.


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