ISSN:
0173-0835
Keywords:
Chemistry
;
Biochemistry and Biotechnology
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Biology
,
Chemistry and Pharmacology
Notes:
A computer-controlled version of recycling isoelectric focusing has been applied to the purification of recombinant human leukocyte interferons (IFNs) from E. coli. The system is based on continuous recycling between a 12-channel heat-exchange reservoir and a 12-channel focusing cell. It contains ten pH meters with probes in each of the flowing channels and two ultraviolet spectrophotometers with five flow cells per instrument. A minicomputer system acquires real-time data of pH and optical density measurements, controls and continuously monitors voltage and power outputs across the focusing cell, and generates on-line multi-color graphics on an x-y plotter. Recycling isoelectric focusing in the presence of 4 M urea resulted in removal of contaminating bacterial pyrogens from recombinant IFN alpha-2 with complete retention of specific antiviral activity. It also produced a 25-fold purification of crude recombinant IFN alpha-5 in a single step and resolved two biologically active species. These data suggest that recycling isoelectric focusing may be incorporated into the design of simple schemes for the purification of a variety of recombinant proteins.
Additional Material:
5 Ill.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/elps.1150071204
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