A flexible approach to automated protein crystallization

Abstract

An automated system for protein crystallization trials is described. Similar to commercial systems in operation, this system is readily assembled at a far lower cost. The software runs on either a microcomputer or a VAX and is both versatile and readily modified for special requirements. The system is capable of pipetting polyethylene glycol solutions accurately. Root-mean-square deviations in the accuracy of pipetted well solutions were similar to results obtained manually. Factors influencing reproducibility are discussed.

Low-resolution X-ray diffraction data obtained from hexagonal crystals of methylamine-treated α2-macroglobulin

“…The desialization was more than 95% complete as measured by the 2-thiobarbituric acid assay (Warren, 1959). Screening for crystallization conditions was performed by vapour diffusion in Cryschem trays (Charles Supper Co., Massachusetts, USA) by the use of a modified Gilson crystallization robot (Oldfield, Ceska & Brady, 1991). Crystallization at 277 K was performed in a cooling incubator.…”

Analysis and crystallization of a 25 kDa C‐terminal fragment of cloned elongation factor Ts fromEscherichia coli

“…Since the reservoir controls the pH when using ammonium sulphate [36] all reservoirs contained 10 mM MES or HEPES and were adjusted to the desired pH with Tris. All crystallizations were set up using a modified Gilson crystallization robot [37]. All solutions used for crystallization were filtered through 0.45 pm pore size filters prior to use.…”

Comparative Studies of Protein Crystallization by Vapour-Diffusion and Microbatch Techniques

“…Cox & Weber, 1987;Ward, Perozzo & Zuk, 1988;Chayen, Shaw Stewart, Maeder & Blow, 1990;Rubin, Talatous & Larson, 1991;Oldfield, Ceska & Brady, 1991;Chayen, Shaw Stewart & Blow, 1992;Soriano & FontecillaCamps, 1993;Sadaoui, Janin & Lewit-Bently, 1994;Chayen, Shaw Stewart & Baldock, 1994). The fundamental difference between the vapourdiffusion and microbatch methods is that diffusion methods are dynamic systems in which conditions are changing throughout the crystallization process and, hence, there is little control over the experiments once trials have been initiated.…”