Integrated strategy for selective expanded bed ion-exchange adsorption and site-specific protein processing using gene fusion technology
2002 (English)In: Journal of Biotechnology, ISSN 0168-1656, E-ISSN 1873-4863, Vol. 96, no 1, 93-102 p.Article in journal (Refereed) Published
The highly charged domain Z(basic) can be used as a fusion partner to enhance adsorption of target proteins to cation exchanging resins at high pH-values. In this paper, we describe a strategy for purification of target proteins fused to Z(basic) at a constant physiological pH using cation exchange chromatography in an expanded bed mode. We show that two proteins, Klenow DNA polymerase and the viral protease 3C, can be efficiently purified from unclarified Escherichia coli homogenates in a single step with a selectivity analogous to what is normally achieved by affinity chromatography. The strategy also includes an integrated site-specific removal of the Z(basic) purification handle to yield a free target protein.
Place, publisher, year, edition, pages
2002. Vol. 96, no 1, 93-102 p.
protein A, Z(basic), ion-exchange chromatography, expanded bed adsorption, DNA-POLYMERASE-I, ESCHERICHIA-COLI, PURIFICATION, RECOVERY, COXSACKIEVIRUS-B3, CHROMATOGRAPHY, EXPRESSION, EFFICIENT, CELLS
Biochemistry and Molecular Biology
IdentifiersURN: urn:nbn:se:kth:diva-13276DOI: 10.1016/S0168-1656(02)00040-8ISI: 000176318300010OAI: oai:DiVA.org:kth-13276DiVA: diva2:323049
QC 201006092010-06-092010-06-092010-10-20Bibliographically approved