Protein Crystallization in a Microfluidic Contactor with Nafion<sup>®</sup>117 Membranes
M. Polino,
H. S. Rho,
M. P. Pina,
R. Mallada,
A. L. Carvalho,
M. J. Romão,
Isabel Coelhoso,
J. G. E. Gardeniers,
J. G. Crespo,
Carla A. M. Portugal
Affiliations
M. Polino
LAQV-REQUIMTE, Department of Chemistry, NOVA School of Science and Technology, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal
H. S. Rho
Department of Instructive Biomaterials Engineering, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, 6229 ER Maastricht, The Netherlands
M. P. Pina
Instituto de Nanociencia y Materiales de Aragón (INMA), Universidad de Zaragoza-CSIC, 50009 Zaragoza, Spain
R. Mallada
Instituto de Nanociencia y Materiales de Aragón (INMA), Universidad de Zaragoza-CSIC, 50009 Zaragoza, Spain
A. L. Carvalho
UCIBIO–Applied Molecular Biosciences Unit, Department of Chemistry, School of Science and Technology, NOVA University Lisbon, 2819-516 Caparica, Portugal
M. J. Romão
UCIBIO–Applied Molecular Biosciences Unit, Department of Chemistry, School of Science and Technology, NOVA University Lisbon, 2819-516 Caparica, Portugal
Isabel Coelhoso
LAQV-REQUIMTE, Department of Chemistry, NOVA School of Science and Technology, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal
J. G. E. Gardeniers
Mesoscale Chemical Systems, TNW Faculty, University of Twente, P.O. Box 217, 7500 AE Enschede, The Netherlands
J. G. Crespo
LAQV-REQUIMTE, Department of Chemistry, NOVA School of Science and Technology, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal
Carla A. M. Portugal
LAQV-REQUIMTE, Department of Chemistry, NOVA School of Science and Technology, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal
Protein crystallization still remains mostly an empirical science, as the production of crystals with the required quality for X-ray analysis is dependent on the intensive screening of the best protein crystallization and crystal’s derivatization conditions. Herein, this demanding step was addressed by the development of a high-throughput and low-budget microfluidic platform consisting of an ion exchange membrane (117 Nafion® membrane) sandwiched between a channel layer (stripping phase compartment) and a wells layer (feed phase compartment) forming 75 independent micro-contactors. This microfluidic device allows for a simultaneous and independent screening of multiple protein crystallization and crystal derivatization conditions, using Hen Egg White Lysozyme (HEWL) as the model protein and Hg2+ as the derivatizing agent. This microdevice offers well-regulated crystallization and subsequent crystal derivatization processes based on the controlled transport of water and ions provided by the 117 Nafion® membrane. Diffusion coefficients of water and the derivatizing agent (Hg2+) were evaluated, showing the positive influence of the protein drop volume on the number of crystals and crystal size. This microfluidic system allowed for crystals with good structural stability and high X-ray diffraction quality and, thus, it is regarded as an efficient tool that may contribute to the enhancement of the proteins’ crystals structural resolution.
