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Updated: Oct 22, 2025

Crystallization of Proteins on Chip by Microdialysis for In Situ X-ray Diffraction Studies
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Protein Crystallization in a Microfluidic Contactor with Nafion®117 Membranes.

M Polino1, H S Rho2, M P Pina3,4,5

  • 1LAQV-REQUIMTE, Department of Chemistry, NOVA School of Science and Technology, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.

Membranes
|August 26, 2021
PubMed
Summary

This study presents a low-budget microfluidic platform for high-throughput protein crystallization screening. The device efficiently screens conditions, yielding high-quality crystals for X-ray analysis.

Keywords:
Nafion® membranemembrane contactorsprotein crystallizationprotein structuresolute diffusion

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Area of Science:

  • Biophysics
  • Crystallography
  • Materials Science

Background:

  • Protein crystallization for X-ray analysis requires extensive screening of conditions.
  • Current methods are empirical and time-consuming.

Purpose of the Study:

  • To develop a high-throughput, low-budget microfluidic platform for protein crystallization and derivatization screening.
  • To improve the quality and resolution of protein crystals for structural analysis.

Main Methods:

  • A microfluidic device was engineered using a Nafion membrane, channel layer, and wells layer.
  • The platform enabled simultaneous screening of 75 independent micro-contactors.
  • Hen Egg White Lysozyme (HEWL) and Hg2+ were used as model protein and derivatizing agent, respectively.

Main Results:

  • The microfluidic system facilitated controlled transport of water and ions via the Nafion membrane.
  • Diffusion coefficients of water and Hg2+ were evaluated.
  • Protein drop volume positively influenced crystal number and size, yielding high-quality crystals.

Conclusions:

  • The developed microfluidic platform is an efficient tool for optimizing protein crystallization and derivatization.
  • This approach can enhance the structural resolution of protein crystals for X-ray diffraction analysis.