|Title:||Single-step affinity purification of toxic and non-toxic proteins on a fluidics platform|
|Authors :||Miao, Jun|
|Published in :||Lab on a Chip|
|Publisher / Ed. Institution :||Royal Society of Chemistry|
|License (according to publishing contract) :||Licence according to publishing contract|
|Type of review:||Peer review (publication)|
|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|
|Appears in Collections:||Publikationen School of Engineering|
Files in This Item:
There are no files associated with this item.
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.