Recombinant protein purification using thermoseparating aqueous two-phase systems
(1999)- Abstract
- The biotechnology industry has grown considerably over the past twenty years, and the revolution within the field of molecular biology has dramatically improved the possibilities of producing proteins and other important biomolecules. The product of interest will be obtained in a multi-component solution and some challenging purification work has to be performed to recover the target molecule in a pure form. The aim of this thesis is to show that an aqueous two-phase system composed of technical grade polymer(s) and recyclable polymers can be used as an efficient primary recovery step for biological material. The polymers studied are either low-cost starch derivatives or thermoseparating copolymers, e.g. ethylene oxide and propylene oxide... (More)
- The biotechnology industry has grown considerably over the past twenty years, and the revolution within the field of molecular biology has dramatically improved the possibilities of producing proteins and other important biomolecules. The product of interest will be obtained in a multi-component solution and some challenging purification work has to be performed to recover the target molecule in a pure form. The aim of this thesis is to show that an aqueous two-phase system composed of technical grade polymer(s) and recyclable polymers can be used as an efficient primary recovery step for biological material. The polymers studied are either low-cost starch derivatives or thermoseparating copolymers, e.g. ethylene oxide and propylene oxide random copolymers (EOPO polymers). Thermoseparating polymers in water solution will upon heating form a two-phase system where one phase is enriched in polymer and the other depleted. The thermoseparating polymer can be recycled and used in a number of extraction systems. Apolipoprotein A-I (Apo A-I) was chosen as target protein in this study. The protein was purified from cell-free E. coli extracts. Systems composed of EOPO/hydroxypropyl starch had high protein capacity and Apo A-I could be efficiently purified while retaining a high yield. Due to the amphiphilic properties of Apo A-I the purification of the protein could be improved by addition of surfactant. Recycling of thermoseparating polymers has been performed, and the recycled polymers have been shown to be just as efficient for protein purification as fresh polymers. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/40062
- author
- Persson, Josefine
- supervisor
- opponent
-
- Prof Kula, Maria-Regina, Heinrich Heine University, Germany
- publishing date
- 1999
- type
- Thesis
- publication status
- published
- subject
- keywords
- polymer recycling., partitioning, apolipoprotein A-I, thermoseparation, bioseparation, Aqueous two-phase systems, polymer, Biology, Biologi
- pages
- 135 pages
- publisher
- Prof. Folke Tjerneld, Dept. of Biochemistry, Lund University,
- defense location
- Chemical Center sal C
- defense date
- 1999-12-10 13:15:00
- external identifiers
-
- other:ISRN: LUNKDL / (NKBK-1059) / 1-135/1999
- language
- English
- LU publication?
- no
- additional info
- Article: Purification of recombinant apolipoprotein A-1Milano expressed in E. coli using aqueous two-phase extraction followed by temperature induced phase separation.Josefine Persson, Lena Nyström, Hans Ageland and Folke TjerneldJournal of Chromatography B, 711 (1998) 97-109. Article: Purification of recombinant and human apolipoprotein A-1 using surfactant micelles in aqueous two-phase systems; recycling of thermoseparating polymer and surfactant with temperature induced phase separation.Josefine Persson, Lena Nyström, Hans Ageland and Folke TjerneldBiotechnology & Bioengineering 65 (1999) 371-381. Article: Polymer recycling in aqueous two-phase extractions using thermoseparating ethylene oxide - propylene oxide copolymers.Josefine Persson, Anita Kaul and Folke Tjerneld(Submitted). Article: Thermoseparating water/polymer system: a novel one-polymer aqueous two-phase system for protein purification.Hans-Olof Johansson, Josefine Persson and Folke TjerneldBiotechnology & Bioengineering (In press). Article: Purification of protein and recycling of polymers in a new aqueous two- phase system using two thermoseparating polymers.Josefine Persson, Hans-Olof Johansson and Folke TjerneldJournal of Chromatography A (In press). Article: Aqueous polymer two-phase systems formed by new thermoseparating polymers.Josefine Persson, Hans-Olof Johansson, Igor Galaev, Bo Mattiasson and Folke TjerneldBioseparation (Submitted).
- id
- 12823aa8-2684-4701-947a-7f2fca2bc14f (old id 40062)
- date added to LUP
- 2016-04-04 10:00:11
- date last changed
- 2018-11-21 20:56:09
@phdthesis{12823aa8-2684-4701-947a-7f2fca2bc14f,
abstract = {{The biotechnology industry has grown considerably over the past twenty years, and the revolution within the field of molecular biology has dramatically improved the possibilities of producing proteins and other important biomolecules. The product of interest will be obtained in a multi-component solution and some challenging purification work has to be performed to recover the target molecule in a pure form. The aim of this thesis is to show that an aqueous two-phase system composed of technical grade polymer(s) and recyclable polymers can be used as an efficient primary recovery step for biological material. The polymers studied are either low-cost starch derivatives or thermoseparating copolymers, e.g. ethylene oxide and propylene oxide random copolymers (EOPO polymers). Thermoseparating polymers in water solution will upon heating form a two-phase system where one phase is enriched in polymer and the other depleted. The thermoseparating polymer can be recycled and used in a number of extraction systems. Apolipoprotein A-I (Apo A-I) was chosen as target protein in this study. The protein was purified from cell-free E. coli extracts. Systems composed of EOPO/hydroxypropyl starch had high protein capacity and Apo A-I could be efficiently purified while retaining a high yield. Due to the amphiphilic properties of Apo A-I the purification of the protein could be improved by addition of surfactant. Recycling of thermoseparating polymers has been performed, and the recycled polymers have been shown to be just as efficient for protein purification as fresh polymers.}},
author = {{Persson, Josefine}},
keywords = {{polymer recycling.; partitioning; apolipoprotein A-I; thermoseparation; bioseparation; Aqueous two-phase systems; polymer; Biology; Biologi}},
language = {{eng}},
publisher = {{Prof. Folke Tjerneld, Dept. of Biochemistry, Lund University,}},
title = {{Recombinant protein purification using thermoseparating aqueous two-phase systems}},
year = {{1999}},
}