Journal
NATURE PROTOCOLS
Volume 7, Issue 2, Pages 364-373Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2011.448
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Funding
- Garvan Institute of Medical Research
- MedImmune
- Australian National Health and Medical Council
- Australian Research Council
- Cancer Institute, NSW
- UK MRC
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Here we describe protocols for the expression of human antibody fragments in Escherichia coli. Antigen-specific clones are identified by soluble fragment ELISA and concentrated by periplasmic preparation. They are then further purified by affinity chromatography. This article provides an overview of expression and purification strategies for human antibody fragments, as well as detailed protocols for the identification of high-affinity binders and for affinity maturation.
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