Determination of protein crystallization kinetics using a regulated evaporation method

Sameer Talreja, Charles F. Zukoski, Paul J A Kenis

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Proteins are difficult to crystallize and the current methods rely on high throughput screening methods, in which hundreds to thousands of solution conditions are screened to locate and optimize the solution conditions where X-ray diffraction quality crystals are formed. We have developed a method that ensures the formation of a solid phase in each experiment and, as a result, each trial yields information on crystal solubility, and crystal nucleation/growth rates. This method relies on evaporating the solvent from a drop at a predetermined rate set by the dimensions of the crystallization platform. Depending on the initial protein and precipitant concentrations and the rate of evaporation, this method yields different solid phases, including amorphous precipitates, gels, showers of small crystals, a few large crystals, or combinations of those. Once solution conditions resulting in good crystals are found, monitoring the crystal size during the experiment provides a way to extract physico-chemical parameters of importance to crystal growth. We use hen egg-white lysozyme as the model protein in this study. The experiments are highly reproducible in number of crystals formed and growth rates. We apply a population balance model to the experimental data to extract the kinetic parameters for crystal nucleation and growth and compare these values with data from literature for lysozyme. The experimental data of number of crystals and growth rate are in excellent agreement with the predictions of the model.

Original languageEnglish (US)
Title of host publication05AIChE
Subtitle of host publication2005 AIChE Annual Meeting and Fall Showcase, Conference Proceedings
Pages8263
Number of pages1
StatePublished - 2005
Externally publishedYes
Event05AIChE: 2005 AIChE Annual Meeting and Fall Showcase - Cincinnati, OH, United States
Duration: Oct 30 2005Nov 4 2005

Other

Other05AIChE: 2005 AIChE Annual Meeting and Fall Showcase
Country/TerritoryUnited States
CityCincinnati, OH
Period10/30/0511/4/05

ASJC Scopus subject areas

  • Engineering(all)

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