A simplified counter diffusion method combined with a 1D simulation program for optimizing crystallization conditions

J Synchrotron Radiat. 2004 Jan 1;11(Pt 1):45-8. doi: 10.1107/s0909049503023446. Epub 2003 Nov 28.

Abstract

We developed a new protein crystallization method has been developed using a simplified counter-diffusion method for optimizing crystallization condition. It is composed of only a single capillary, the gel in the silicon tube and the screw-top test tube, which are readily available in the laboratory. The one capillary can continuously scan a wide range of crystallization conditions (combination of the concentrations of the precipitant and the protein) unless crystallization occurs, which means that it corresponds to many drops in the vapor-diffusion method. The amount of the precipitant and the protein solutions can be much less than in conventional methods. In this study, lysozyme and alpha-amylase were used as model proteins for demonstrating the efficiency of this method. In addition, one-dimensional (1-D) simulations of the crystal growth were performed based on the 1-D diffusion model. The optimized conditions can be applied to the initial crystallization conditions for both other counter-diffusion methods with the Granada Crystallization Box (GCB) and for the vapor-diffusion method after some modification.

Publication types

  • Comparative Study
  • Evaluation Study
  • Validation Study

MeSH terms

  • Capillary Action
  • Computer Simulation
  • Crystallization / instrumentation
  • Crystallization / methods*
  • Diffusion
  • Models, Chemical*
  • Muramidase / analysis*
  • Muramidase / chemical synthesis
  • Muramidase / chemistry*
  • Quality Control
  • Sodium Chloride / chemistry*
  • alpha-Amylases / analysis*
  • alpha-Amylases / chemical synthesis
  • alpha-Amylases / chemistry*

Substances

  • Sodium Chloride
  • hen egg lysozyme
  • alpha-Amylases
  • Muramidase