High-throughput screens for postgenomics: studies of protein crystallization using microsystems technology.
This paper describes the fabrication of a micromachined miniaturized array of chambers in a 2-mm-thick single crystal (100) silicon substrate for the combinatorial screening of the conditions required for protein crystallization screening (including both temperature and the concentration of crystallization agent). The device was fabricated using standard photolithography techniques, reactive ion etching (RIE) and anisotropic silicon wet etching to produce an array of 10 x 10 microchambers, with each element having a volume of 5 microL. A custom-built temperature controller was used to drive two peltier elements in order to maintain a temperature gradient (between 12 and 40 degrees C) across the device. The performance of the microsystem was illustrated by studying the crystallization of a model protein, hen egg white lysozyme. The crystals obtained were studied using X-ray diffraction at room temperature and exhibited 1.78 A resolution. The problems of delivering a robust crystallization protocol, including issues of device fabrication, delivery of a reproducible temperature gradient, and overcoming evaporation are described.
Juárez-Martínez
G
G
Department of Electronics, University of Glasgow, UK.
Steinmann
P
P
Roszak
A W
AW
Isaacs
N W
NW
Cooper
J M
JM
D003460:Crystallization; D009566:Nitrates; D011506:Proteins / Q000737:chemistry*; D012825:Silicon; D013696:Temperature
D009566:Nitrates; D011506:Proteins; C031618:sodium nitrate; D012825:Silicon