Related Experiment Videos
High-throughput screens for postgenomics: studies of protein crystallization using microsystems technology
G Juárez-Martínez1, P Steinmann, A W Roszak
1Department of Electronics, University of Glasgow, UK.
Analytical Chemistry
|July 26, 2002
Summary
This study presents a novel microfabricated silicon device for protein crystallization screening. The miniaturized array of microchambers enables combinatorial screening of temperature and agent concentration, optimizing crystal growth conditions.
Area of Science:
- Materials Science
- Biotechnology
- Crystallography
Background:
- Protein crystallization is crucial for structural biology, but screening conditions is time-consuming.
- Miniaturization offers potential for high-throughput screening and reduced sample consumption.
Purpose of the Study:
- To fabricate and characterize a micromachined silicon device for combinatorial protein crystallization screening.
- To optimize protein crystallization conditions by systematically varying temperature and crystallization agent concentration.
Main Methods:
- Fabrication using photolithography, reactive ion etching (RIE), and anisotropic silicon wet etching.
- Creation of a 10x10 array of 5 microL microchambers on a silicon substrate.
- Implementation of a temperature gradient (12-40°C) using a custom controller and Peltier elements.
Main Results:
- Successful fabrication of a miniaturized protein crystallization screening device.
- Demonstration of the device's performance using hen egg white lysozyme.
- Obtained protein crystals diffracted X-rays to 1.78 Å resolution.
Conclusions:
- The developed microsystem is effective for combinatorial screening of protein crystallization conditions.
- Challenges in device fabrication, temperature control, and evaporation mitigation were addressed.
- This technology facilitates robust crystallization protocol development for structural studies.