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Characterization of Glycoproteins with the Immunoglobulin Fold by X-Ray Crystallography and Biophysical Techniques
Published on: July 5, 2018
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Large-scale crystallization of proteins for purification and formulation
1Institute of Biochemical Engineering, Technische Universität München, Boltzmannstr. 15, 85748, Garching, Germany, hekmat@lrz.tum.de.
Bioprocess and Biosystems Engineering
|February 22, 2015
Summary
Protein crystallization, a long-standing technique, is still relevant for purifying and formulating therapeutic proteins. Advances in understanding its mechanisms and scalability offer new platform purification possibilities.
Area of Science:
- Biochemistry
- Chemical Engineering
- Crystallography
Background:
- Protein crystallization has been utilized for approximately 170 years.
- Early recognition of salt effects (dehydrating, kosmotropic/chaotropic) and organic solvents.
- Despite extensive research, fundamental understanding of protein solvation and interactions remains incomplete.
Purpose of the Study:
- To explore preparative protein crystallization techniques.
- To demonstrate protein crystallization as a viable alternative to chromatography.
- To investigate protein crystallization for therapeutic protein formulation.
Main Methods:
- Case studies illustrating preparative protein crystallization.
- Application of pre-treatment procedures (precipitation, extraction) for impure solutions.
- Utilizing maximum local energy dissipation for process scale-up.
Main Results:
- Demonstrated feasibility of preparative crystallization from impure protein solutions.
- Protein crystallization can effectively replace multiple chromatography steps.
- Crystalline proteins offer enhanced purity and highly concentrated therapeutic doses.
- Scalability of crystallization processes was successfully shown.
Conclusions:
- Protein crystallization requires case-specific process design due to its variable nature.
- Crystallization serves as an attractive formulation strategy for therapeutic proteins.
- Future integration into platform purification processes is anticipated with molecular modeling and protein engineering.
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