Purification of phage display-modified bacteriophage T4 by affinity chromatography
- نوع فایل : کتاب
- زبان : انگلیسی
- مؤلف : Anna Oślizło, Paulina Miernikiewicz, Agnieszka Piotrowicz, Barbara Owczarek, Agnieszka Kopciuch, Grzegorz Figura, Krystyna Dąbrowska*
- چاپ و سال / کشور: 2011
Description
Affinity chromatography is one of the most efficient protein purification strategies. This technique comprises a one-step procedure with a purification level in the order of several thousand-fold, adaptable for various proteins, differentiated in their size, shape, charge, and other properties. The aim of this work was to verify the possibility of applying affinity chromatography in bacteriophage purification, with the perspective of therapeutic purposes. T4 is a large, icosahedral phage that may serve as an efficient display platform for foreign peptides or proteins. Here we propose a new method of T4 phage purification by affinity chromatography after its modification with affinity tags (GST and Histag) by in vivo phage display. As any permanent introduction of extraneous DNA into a phage genome is strongly unfavourable for medical purposes, integration of foreign motifs with the phage genome was not applied. The phage was propagated in bacteria expressing fusions of the phage protein Hoc with affinity tags from bacterial plasmids, independently from the phage expression system.
