Title: Single-step affinity purification of toxic and non-toxic proteins on a fluidics platform
Authors : Miao, Jun
Wu, Wei
Spielmann, Thomas
Belfort, Marlene
Derbyshire, Victoria
Belfort, Georges
Published in : Lab on a Chip
Volume(Issue) : 2005
Issue : 3
Pages : 248
Pages to: 253
Publisher / Ed. Institution : Royal Society of Chemistry
Issue Date: 2005
License (according to publishing contract) : Licence according to publishing contract
Type of review: Peer review (publication)
Language : English
Subjects : Chromatography, Affinity; Endodeoxyribonucleases; Escherichia coli; Gene Expression Regulation, Bacterial; Hydrogen-Ion Concentration; Inteins; Microfluidics; Protein Splicing; Proteomics; Recombinant Fusion Proteins; Time Factors
Subject (DDC) : 572: Biochemistry
Abstract: Single-step fusion-based affinity purification of proteins with pH-controllable linkers was carried out in a fluidic device. The linkers were previously derived from self-splicing protein elements called inteins. Two different linkers were generated to solve two distinct separation problems: one for rapid single-step affinity purification of a wide range of proteins, and the other specifically for the purification of cytotoxic proteins. Scale-down factors of 185 resulted in separations in a 27 microl bed-volume. A rotating CD format was chosen because of its simplicity in effecting fluid movement through centrifugal force without the complications associated with electro-osmosis and other pumping methods. The design and fabrication of the fluidic device and the protein purification process are described. This work, which demonstrates the purification of active proteins by two distinct fluidic separations, is widely applicable to small-scale massively parallel proteomic separations.
Departement: School of Engineering
Organisational Unit: Institute of Materials and Process Engineering (IMPE)
Publication type: Article in scientific journal
DOI : 10.1039/b413292k
ISSN: 1473-0197
URI: https://digitalcollection.zhaw.ch/handle/11475/16248
Appears in Collections:Publikationen School of Engineering

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