Extension of the selection of protein chromatography and the rate model to affinity chromatographyy
Author
dc.contributor.author
Sandoval, G.
Author
dc.contributor.author
Shene, C.
es_CL
Author
dc.contributor.author
Andrews Farrow, Bárbara
es_CL
Author
dc.contributor.author
Asenjo de Leuze, Juan
es_CL
Admission date
dc.date.accessioned
2014-01-07T13:30:33Z
Available date
dc.date.available
2014-01-07T13:30:33Z
Publication date
dc.date.issued
2010-08-03
Cita de ítem
dc.identifier.citation
J. Mol. Recognit. 2010; 23: 609–617
en_US
Identifier
dc.identifier.uri
https://repositorio.uchile.cl/handle/2250/125990
General note
dc.description
Artículo de publicación ISI
en_US
Abstract
dc.description.abstract
The rational selection of optimal protein purification sequences, as well as mathematical models that simulate and
allow optimization of chromatographic protein purification processes have been developed for purification procedures
such as ion-exchange, hydrophobic interaction and gel filtration chromatography. This paper investigates the
extension of such analysis to affinity chromatography both in the selection of chromatographic processes and in the
use of the rate model for mathematical modelling and simulation. Two affinity systems were used: Blue Sepharose and
Protein A. The extension of the theory developed previously for ion-exchange and HIC chromatography to affinity
separations is analyzed in this paper. For the selection of operations two algorithms are used. In the first, the value of
h, which corresponds to the efficiency (resolution) of the actual chromatography and, S, which determines the
amount of a particular contaminant eliminated after each separation step, which determines the purity, have to be
determined. It was found that the value of both these parameters is not generic for affinity separations but will
depend on the type of affinity system used and will have to be determined on a case by case basis. With Blue
Sepharose a salt gradient was used and with Protein A, a pH gradient. Parameters were determined with individual
proteins and simulations of the protein mixtures were done. This approach allows investigation of chromatographic
protein purification in a holistic manner that includes ion-exchange, HIC, gel filtration and affinity separations for the
first time.