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J Biotechnol. 2012 Jan 20;157(2):315-9. doi: 10.1016/j.jbiotec.2011.09.028. Epub 2011 Oct 5.

Protein precipitation by polyethylene glycol: a generalized model based on hydrodynamic radius.

Author information

1
Bioprocessing Technology Institute, Agency for Science, Technology and Research, Centros #06-01, Singapore 138668. siowleng.sim@a-bio.com

Abstract

PEGs for protein precipitation are usually classified by molecular weight. The higher molecular weight precipitants are more efficient but result in higher viscosity. Following empirical evidence that the precipitation efficiency is more comprehensively characterized by PEG hydrodynamic radius (r(h,PEG)) than molecular weight, this paper proposes a model to explicate the significance of r(h,PEG). A general expression was formulated to characterize the PEG effect exclusively by r(h,PEG). The coefficients of a linearized form were then fitted using empirical solubility data. The result is a simple numerical relation that models the efficiency of general-shaped PEG precipitants as a function of r(h,PEG) and protein hydrodynamic radius (r(h,prot)). This equation also explains the effects of environmental conditions and PEG branching. While predictions by the proposed correlation agree reasonably well with independent solubility data, its simplicity gives rise to potential quantitative deviations when involving small proteins, large proteins and protein mixtures. Nonetheless, the model offers a new insight into the precipitation mechanism by clarifying the significance of r(h,PEG). This in turn helps to refine the selection criterion for PEG precipitants.

PMID:
22001847
DOI:
10.1016/j.jbiotec.2011.09.028
[Indexed for MEDLINE]

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