Generation of human single domain antibody repertoires by kunkel mutagenesis

Abstract

Human antibody single domains are a promising new class of antibody fragments. Here we describe methods for the cloning of human VH and VL genes into phage and phagemid vectors. Furthermore, we provide detailed protocols for the generation of single domain antibody libraries by Kunkel mutagenesis and the analysis of diversity by DNA sequencing and superantigen binding. © 2012 Springer Science+Business Media, LLC.