Protein Crystallization in a Microfluidic Contactor with Nafion®117 Membranes

Polino, M. ; Rho, H. S. ; Pina, M. P. (Universidad de Zaragoza) ; Mallada, R. (Universidad de Zaragoza) ; Carvalho, A. L. ; Romão, M. J. ; Coelhoso, Isabel ; Gardeniers, J. G. E. ; Crespo, J. G. ; Portugal, Carla A. M.

Resumen: 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.

Idioma: Inglés

DOI: 10.3390/membranes11080549

Año: 2021

Publicado en: Membranes 11, 8 (2021), 549 [18 pp.]
ISSN: 2077-0375

Factor impacto JCR: 4.562 (2021)

Categ. JCR: POLYMER SCIENCE rank: 21 / 90 = 0.233 (2021) - Q1 - T1
Categ. JCR: MATERIALS SCIENCE, MULTIDISCIPLINARY rank: 132 / 345 = 0.383 (2021) - Q2 - T2
Categ. JCR: CHEMISTRY, PHYSICAL rank: 69 / 165 = 0.418 (2021) - Q2 - T2
Categ. JCR: ENGINEERING, CHEMICAL rank: 47 / 143 = 0.329 (2021) - Q2 - T1

Factor impacto CITESCORE: 3.7 - Chemical Engineering (Q2)

Factor impacto SCIMAGO: 0.517 - Process Chemistry and Technology (Q2) - Chemical Engineering (miscellaneous) (Q2)

Tipo y forma: Article (Published version)

Área (Departamento): Área Ingeniería Química (Dpto. Ing.Quím.Tecnol.Med.Amb.)