alexa Mathematical Modelling of Protein Precipitation Based on the Phase Equilibrium for an Antibody Fragment from E. coli Lysis
ISSN: 2155-9821

Journal of Bioprocessing & Biotechniques
Open Access

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Research Article

Mathematical Modelling of Protein Precipitation Based on the Phase Equilibrium for an Antibody Fragment from E. coli Lysis

Yu Ji and Yuhong Zhou*
Department of Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, UK
Corresponding Author : Yuhong Zhou
University College London
Department of Biochemical Engineering
Torrington Place, London WC1E 7JE, UK
Tel: +44(0)207 679 3815
Fax: +44 (0)207 916 3943
E-mail: [email protected]
Received June 06, 2013; Accepted July 21, 2013; Published July 26, 2013
Citation: Ji Y, Zhou Y (2013) Mathematical Modelling of Protein Precipitation Based on the Phase Equilibrium for an Antibody Fragment from E. coli Lysis. J Bioprocess Biotech 3:129 doi: 10.4172/2155-9821.1000129
Copyright: © 2013 Ji Y, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
 

Abstract

Precipitation is an important operation in biopharmaceutical purification yet the mechanism of protein precipitation in multi-component solutions is not well understood. Existing models lack fundamental understanding of the process. In this paper, a new model describing how the protein solubility changes in the protein precipitation is proposed and is based on the phase equilibrium of the light liquid phase and dense solid phase. The model structure is generic and robust. It adequately reflects the non-linearity of protein precipitation kinetics and thus provides new fundamental insights into the protein precipitation in multi-component, complex protein solution.
Two feed stocks of a pure fragment antigen-binding (Fab’) solution obtained by chromatographic purification and a clarified Fab’ homogenate solution from E. coli were used to examine the effect of ammonium sulphate concentrations and pH conditions on precipitation. It was found that the model can describe pure Fab’ precipitation well, and identify the non-ideal behavior of Fab’ precipitation in multi-component homogenates. Through statistical analysis, the model parameters have been further reduced from 8 to 4. The quality of the model is such that errors were within the acceptable statistical confidence limits, even when applied to multi-component impurity precipitation. The new model with fewer parameters is better than existing empirical models in reflecting the salting-in and salting-out effect of the protein precipitation. This demonstrated that the structure of the model is sound and over-fitting in the parameter estimation is avoided. The model can be applied directly to industrial processes for protein precipitation process design after appropriate calibration with the required operating conditions of pH and salt concentration.

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