title: A two-phagemid system for the creation of non-phage displayed antibody libraries approaching one trillion members.
abstract: We have designed a two-phagemid system for the construction of very large non-phage displayed Fab antibody libraries in E. coli approaching 10(12) members. The system can accommodate both periplasmic and cytoplasmic Fab expression and should prove useful for the direct selection of functional antibodies by genetic techniques. We successfully alleviate problems of Fab vector instability and report a set of improved 5' primers for the amplification of mouse Ig V(H)95% of mouse Ig V(H) genes and minimize the amount of N-terminal amino acid changes while maintaining the flexibility of periplasmic or cytoplasmic antibody expression in E. coli.
authors.affiliation: The Pennsylvania State University, Department of Chemistry, 414 Wartik Laboratory, University Park, PA 16802, USA.
authors.forename: M
authors.initials: M
authors.lastname: Ostermeier
authors.forename: S J
authors.initials: SJ
authors.lastname: Benkovic
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000235:genetics*; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005803:Genes, Immunoglobulin; D005822:Genetic Vectors; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D051379:Mice; D019151:Peptide Library; D010957:Plasmids / Q000235:genetics; D020798:Two-Hybrid System Techniques
chemical_list: D000906:Antibodies; D017931:DNA Primers; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D019151:Peptide Library