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Large-scale crystallization of proteins for purification and formulation

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Abstract

Since about 170 years, salts were used to create supersaturated solutions and crystallize proteins. The dehydrating effect of salts as well as their kosmotropic or chaotropic character was revealed. Even the suitability of organic solvents for crystallization was already recognized. Interestingly, what was performed during the early times is still practiced today. A lot of effort was put into understanding the underlying physico-chemical interaction mechanisms leading to protein crystallization. However, it was understood that already the solvation of proteins is a highly complex process not to mention the intricate interrelation of electrostatic and hydrophobic interactions taking place. Although many basic questions are still unanswered, preparative protein crystallization was attempted as illustrated in the presented case studies. Due to the highly variable nature of crystallization, individual design of the crystallization process is needed in every single case. It was shown that preparative crystallization from impure protein solutions as a capture step is possible after applying adequate pre-treatment procedures like precipitation or extraction. Protein crystallization can replace one or more chromatography steps. It was further shown that crystallization can serve as an attractive alternative means for formulation of therapeutic proteins. Crystalline proteins can offer enhanced purity and enable highly concentrated doses of the active ingredient. Easy scalability of the proposed protein crystallization processes was shown using the maximum local energy dissipation as a suitable scale-up criterion. Molecular modeling and target-oriented protein engineering may allow protein crystallization to become part of a platform purification process in the near future.

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Acknowledgments

The author thanks Prof. Dirk Weuster-Botz, Head of the Institute of Biochemical Engineering, Technische Universität München, Germany, for the excellent support of this work and for the opportunity of using the outstanding infrastructure at the Institute of Biochemical Engineering. The author also thanks Dirk Hebel and Benjamin Smejkal for performing the work presented in some of the case studies. The funding of the Federal Ministry of Education and Research, Germany (Grant No. 0315335B), and Novartis Pharma AG, Switzerland, is gratefully acknowledged.

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    Dariusch Hekmat

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Hekmat, D. Large-scale crystallization of proteins for purification and formulation. Bioprocess Biosyst Eng 38, 1209–1231 (2015). https://doi.org/10.1007/s00449-015-1374-y

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