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Expression of high affinity human antibody fragments in bacteria

Abstract

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.

Type Journal
Authors Rouet, R.; Lowe, D.; Dudgeon, K.; Roome, B.; Schofield, P.; Langley, D.; Andrews, J.; Whitfeld, P.; Jermutus, L.; Christ, D.:
Garvan Authors A/Prof Daniel Christ , Dr David Langley , Dr Peter Schofield , Dr Peter Whitfeld , Romain Rouet
Publisher Name NAT PROTOC
Published Date 2012-02-02 00:00:00
Published Volume 7
Published Issue 2
Published Pages 364-73
URL http://www.ncbi.nlm.nih.gov/pubmed/22301775
Status Published In-print
OpenAccess Link https://publications.gimr.garvan.org.au/download.php?11201_12108/12_Rouet_NatProt.pdf