Single-step affinity purification of toxic and non-toxic proteins on a fluidics platform

Miao, Jun; Wu, Wei; Spielmann, Thomas; Belfort, Marlene; Derbyshire, Victoria; Belfort, Georges

doi:10.1039/b413292k

Publication type:	Article in scientific journal
Type of review:	Peer review (publication)
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
DOI:	10.1039/b413292k
Published in:	Lab on a Chip
Volume(Issue):	2005
Issue:	3
Page(s):	248
Pages to:	253
Issue Date:	2005
Publisher / Ed. Institution:	Royal Society of Chemistry
ISSN:	1473-0197 1473-0189
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.
URI:	https://digitalcollection.zhaw.ch/handle/11475/16248
Fulltext version:	Published version
License (according to publishing contract):	Licence according to publishing contract
Departement:	School of Engineering
Organisational Unit:	Institute of Materials and Process Engineering (IMPE)
Appears in collections:	Publikationen School of Engineering

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Miao, J., Wu, W., Spielmann, T., Belfort, M., Derbyshire, V., & Belfort, G. (2005). Single-step affinity purification of toxic and non-toxic proteins on a fluidics platform. Lab on a Chip, 2005(3), 248–253. https://doi.org/10.1039/b413292k

Miao, J. et al. (2005) ‘Single-step affinity purification of toxic and non-toxic proteins on a fluidics platform’, Lab on a Chip, 2005(3), pp. 248–253. Available at: https://doi.org/10.1039/b413292k.

J. Miao, W. Wu, T. Spielmann, M. Belfort, V. Derbyshire, and G. Belfort, “Single-step affinity purification of toxic and non-toxic proteins on a fluidics platform,” Lab on a Chip, vol. 2005, no. 3, pp. 248–253, 2005, doi: 10.1039/b413292k.

MIAO, Jun, Wei WU, Thomas SPIELMANN, Marlene BELFORT, Victoria DERBYSHIRE und Georges BELFORT, 2005. Single-step affinity purification of toxic and non-toxic proteins on a fluidics platform. Lab on a Chip. 2005. Bd. 2005, Nr. 3, S. 248–253. DOI 10.1039/b413292k

Miao, Jun, Wei Wu, Thomas Spielmann, Marlene Belfort, Victoria Derbyshire, and Georges Belfort. 2005. “Single-Step Affinity Purification of Toxic and Non-Toxic Proteins on a Fluidics Platform.” Lab on a Chip 2005 (3): 248–53. https://doi.org/10.1039/b413292k.

Miao, Jun, et al. “Single-Step Affinity Purification of Toxic and Non-Toxic Proteins on a Fluidics Platform.” Lab on a Chip, vol. 2005, no. 3, 2005, pp. 248–53, https://doi.org/10.1039/b413292k.