Purification and Formulation of a Therapeutic Antibody by Crystallization

Benjamin Smejkal, doctoral thesis Technische Universität München, 2013

High costs for purification of biopharmaceuticals may be reduced by alternative process steps. Hence, protein crystallization on a technical scale has been investigated in this work. For stirred tanks, the maximum local energy dissipation was identified as scale-up criterion. Using an optimized power input at appropriate crystallization conditions, a crystal yield of 98 % was already achieved within a few minutes for the investigated therapeutic antibody on a L-scale. Furthermore, protein crystallization was enabled directly from pretreated crude fermentation broth by the removal of salts inhibiting nucleation. Thus, a new purification process was established without the need for expensive chromatography steps. Finally, it was shown that antibody crystals with an adjusted crystal size could be used for stable storage and efficient injection.

 

Publications

• Hekmat D, Huber M, Lohse C, von den Eichen N, Weuster-Botz D (2017): Continuous crystallization of proteins in a stirred classified product removal tank with a tubular reactor in bypass. Cryst Growth Des, DOI: 10.1021/acs.cgd.7b00436.
• Smejkal B, Agrawal NJ, Helk B, Schulz H, Giffard M, Mechelke M, Ortner F, Heckmeier P, Trout BL, Hekmat D (2013): Fast and scalable purification of a therapeutic full-length antibody based on process crystallization. Biotechnol Bioeng 110: 2452-2461.
• Smejkal B, Helk B, Rondeau JM, Anton S, Wilke A, Scheyerer P, Fries J, Hekmat D, Weuster-Botz D (2013): Protein crystallization in stirred systems – Scale-up via the maximum local energy dissipation. Biotechnol Bioeng 110: 1956-1963.