A microfluidic device for both on-chip dialysis protein crystallization and in situ X-ray diffraction

Abstract

This paper reports a versatile microfluidic chip developed for on-chip crystallization of proteins through the dialysis method and in situ X-ray diffraction experiments.

Easy-to-Use Osmosis-Based Microfluidic Chip for Protein Crystallization: Application to a Monoclonal Antibody

“…Among these microuidic developments, we can notably mention batch crystallization in closed microchannels or droplets of a few nanoliters, 1322 controlled interdiusion of proteins and precipitants, 2326 or chips integrating membranes to mimic dialysis crystallization. 27,28 Beyond the screening of crystallization conditions for structural analysis (sometimes in situ), some of these devices have also been developed for the acquisition of data dicult to measure on a macroscopic scale (phase diagram and solubility, 14,22,23 nucleation kinetics, 1820 protein-protein interactions, 29 etc. ).…”

Crystallization of Proteins on Chip by Microdialysis for <i>In Situ</i> X-ray Diffraction Studies

“…High throughput screening and optimization of crystallization conditions can be achieved 30 , 31 in well-based 32 , droplet-based 33 , or valve-actuated 34 devices. In situ X-ray diffraction experiments of challenging protein targets at room temperature have been conducted in microchips fabricated from various materials such as PDMS (polydimethylsiloxane), COC (cyclic olefin copolymer), PMMA (poly(methyl methacrylate)) 21 , 22 , 26 , 28 , 29 , graphene films 23 , Kapton 35 , epoxy glue 6 , or NOA (Norland Optical Adhesive) 19 and the materials' transparency to X-ray radiation and their contribution to background noise have been evaluated. Moreover, microchips have been designed to couple the in situ and the serial data collection strategies in a single tool for X-ray protein crystallography experiments at synchrotron sources 23 , 35 , 36 and XFELs 7 .…”

“…In this work, a protocol is described for producing dialysis microchips incorporating regenerated cellulose (RC) dialysis membranes of different MWCO in order to perform crystallization assays on-chip and in situ X-ray diffraction data collection. The materials comprising the microchips have been evaluated for their transparency to X-rays 19 and the devices can be set directly in front of the X-ray beam for room temperature in situ diffraction experiments, excluding the manual handling and minimizing the degradation of fragile protein crystals. In a case study, hen egg-white lysozyme crystals were grown onchip via microdialysis generating a uniformly sized population.…”

Microfluidic dialysis using photo-patterned hydrogel membranes in PDMS chips

“…In particular, nanoporous membranes are attracting a great deal of interest due to the many applications in the field of dialysis and ultra-filtration. Many works integrate a commercial dialysis membrane by sandwiching it between two microfluidic chips, with applications as diverse as the study of phase diagrams of colloidal systems [8], protein crystallization [9], or to investigate osmotically-driven flows [10] and fouling in ultra-filtration processes [11]. But it is in the field of analytical biochemistry that this type of technique is of particular interest with numerous applications such as on-chip microdialysis sample cleanup, fractionation, concentration, separation, etc, see e.g.…”