Tailored Hydrogel Membranes for Efficient Protein Crystallization

• Published in: Advanced functional materials 2014-03, Vol.24 (11), p.1582-1590
• Main Authors: Profio, Gianluca Di, Polino, Mariella, Nicoletta, Fiore P., Belviso, Benny D., Caliandro, Rocco, Fontananova, Enrica, Filpo, Giovanni De, Curcio, Efrem, Drioli, Enrico
• Format: Article
• Language: English
• Subjects: composite membranes; Concentration (composition); crystal engineering; Crystallization; Crystals; Diffraction; gels; Hydrogels; Membranes; Nanostructure; protein crystallization; Proteins; X-ray diffraction
• ISSN: 1616-301X; 1616-3028
• DOI: 10.1002/adfm.201302240

Crystallization still represents the bottleneck in the process of protein structure determination at high resolution, despite high‐throughput structural genomics programs requiring optimized crystallization strategies regarding crystal quality, time, success rate, reproducibility, and used protein amount. On the other hand, the development of suitable materials for controlled heterogeneous nucleation might facilitate biomacromolecular crystallization in a variety of experimental conditions which are not conventionally fruitful. Here, the possibility to fabricate hydrogel membranes displaying controlled chemical composition and nanostructure and to use them as heterogeneous supports for biomacromolecular crystallization is demonstrated. Diverse gel morphologies are obtained by controlling phase separation kinetics during gel layer formation on membrane support. These composite materials are found to increase the efficiency of the crystallization process so that crystals with enhanced diffraction properties are produced at lower protein concentration than conventional technique, thus affording the possibility to improve current approaches to protein crystallization and to be adapted to specific targets. Composite membranes supporting a homogeneous hydrogel layer, with tailored chemical composition, nano‐architecture (mesh size), and selected morphologies, are fabricated and tested for protein crystallization. As such, the general advantages of gel and membrane‐assisted crystallization are combined to produce crystals displaying enhanced diffraction features at lower protein concentration with respect to conventionally grown crystals.