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Construction of a Human Antibody Domain (VH) Library

), called a “domain antibody”, or “dAb”, is approximately four times smaller than a Fab and half the size of a scFv. Here, we describe the construction of a phage-displayed VH library and an approach to introduce genetic diversity in this library, where both diverse

Expression and Isolation of Recombinant Antibody Fragments in E. coli

linker into expression constructs to produce scFv. Further variants include the addition of a range of suitable (C-terminal) tags for the purification or detection of expressed protein, such as a human Cκ domain (scAb) (2 ), c-myc, or hexahistidine.

Expression, Purification, and Characterization of Engineered Antibody CH2 and VH Domains

scaffolds of much smaller size and high stability. Immunoglobulin-based scaffolds including Fab (∼50 kD), ScFv (∼30 kD), and VH domain (termed domain antibody, dAb) (∼15 kD) have been well established. Recently, a new scaffold based on human IgG1 CH2 domain