Lisa J. Garrard

Selection of an anti-IGF-1 Fab from a Fab phage library created by mutagenesis of multiple CDR loops

Diverse Fab libraries containing 2-3 x 10(8) members were generated by randomizing amino acid residues within four of the six complementarity determining regions of a humanized version of an anti-HER-2 Ab (hu4D5). These libraries were subsequently displayed on the surface of the filamentous bacteriophage M13 and selected for binding to three proteins: CD4, insulin-like growth factor 1 (IGF-1), and tissue plasminogen activator. An Fab-bacteriophage was isolated that showed specific binding to IGF-1. The affinity of this Fab was determined to be 3.5 microM.

Antibody engineering using very long template-assembled oligonucleotides

We have previously constructed single-stranded DNA fragments up to 361 nucleotides long by the ligation of shorter synthetic fragments assembled on a closely homologous template and used them for site-directed mutagenesis. Here we have extended the utility of such long oligonucleotides for protein engineering in two ways. First oligonucleotides have been assembled on a template to which they show only relatively weak sequence homology (

Antibody expression in bacteriophage systems: The future of monoclonal antibodies?

Bacteriophage systems have been utilized to express and isolate antibodies. This promising technology has been evolving rapidly and has the potential to revolutionize the way in which monoclonal antibodies are generated. This review focuses on the many recent advances that have been made in obtaining monoclonal antibodies from bacteriophage systems.