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Design of Aqueous Two-Phase Protein Extraction Process

Ahmad, Murni (2006) Design of Aqueous Two-Phase Protein Extraction Process. PhD thesis, Universiti Teknologi Petronas.

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Abstract

Aqueous Two-Phase Extraction Systems (ATPES) comprise a liquid-liquid extraction technique that exploits phase separation phenomenon to recover protein. The two-liquid-phase systems are formed when two phase forming agent,s are aclclecl together in certain proportions or conditions that exceed a critical, and cu·e under a ma.ximum concentration, temperature or pH threshold. In this water-rich system, a protein selectively partitions into one of the phases, and this selectivity can be enhanced. Other than offering high yield for protein recovery, ATPES also allows continuous steady-state operation with high capacity, easy engineering scale-up, concentration of target species, and most encouragingly, it can directly extract product from crude feedstock rnaking integration with upstream processing possible. Despite the fact that ATPES has the potential to fulfill the dynamic and evolving clernand by indus try as an efficient, huge-scale; primary downstream process to extract protein, there are still a limited number of applications at industrial scale. This is clue to the economic concerns regarding the arnount of polymer required to extract a small amount of protein as some of the polymers are expensive. Furthermore, there are limited design methods available compared to other separation techniques used in chemical process industries. In this thesis, we present a heuristic design approach to investigate ATPES as a protein extraction technique. This work involves developing a thermodynamic modeling framework that uses the Flory-Huggins model [52, 77] to calculate the liquid-liquid equilibrium behavior in aqueous polyrner mixture and polymer-salt ATPESs. This thermodynamic work, based on the minimization of the Gibbs energy of mixing, is able to represent the ATPES equilibrium behavior in the presence of a target protein, a contaminant and other phaseforming components in dilute concentrations within a design range. This work is extended to the evaluation of the protein partitioning behavior in these systems. Our work also focuses on the development of an efficient Aowsheet simulation and evaluation framework. \Ve sirnulate the flowsheets for a.n extraction unit with one and two stages that contain ATPESs. \Ve address a. feed feasibility issue encountered in two-stage configurations by using a heuristic approach. This flowsheet simulation framework integrates the therrnodynmnic framework, heuristic feasible feed calculations and an iterative recycle convergence. '\'e develop a cost minimization of the flowsheets subject to the fe<lSible two-phase region and the kinetics of the ATPES in the flowsheet, to obtain the optimal design. In the last step of our computational work, we describe aJ1 approach to designing cost-optimal flowsheets, that balances the requirements for yield, purity, settling kinetics and the amount of phase-forming components used. 'Ve demonstrate the application of our work on calculations for a model system comprising phosphofructokinase and ovalbumin. In addition, an experimental model system that is based on the extraction of recombinant green fluorescent protein using polymer mixture and polymer-salt ATPESs, is used to test the applicability of our framework. 'Ve fit the empirical liquid-liquid equilibrium data to obtain Flory-Huggins model [52, 77] pararneters and demonstrate the initial feasibility of the framework to design one-stage flowsheet for the protein's ATPES-basecl extraction. "'e also show the possible implementation of our approach to design a two-stage extraction unit. '\'e discuss some issues that arose in our work and suggest future work to extend the applicability of this work.

Item Type: Thesis (PhD)
Academic Subject : Academic Department - Chemical Engineering - Advance Process Control
Subject: T Technology > TP Chemical technology
Divisions: Engineering > Chemical
Depositing User: Users 2053 not found.
Date Deposited: 30 Sep 2013 16:55
Last Modified: 25 Jan 2017 09:46
URI: http://utpedia.utp.edu.my/id/eprint/8049

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