title: Platform for high-throughput antibody selection using synthetically-designed antibody libraries.
abstract: Synthetic humanized antibody libraries are frequently generated by random incorporation of changes at multiple positions in the antibody hypervariable regions. Although these libraries have very large theoretical diversities (>10(20)), the practical diversity that can be achieved by transformation of Escherichia coli is limited to about 10(10). To constrain the practical diversity to sequences that more closely mimic the diversity of natural human antibodies, we generated a scFv phage library using entirely pre-defined complementarity determining regions (CDR). We have used this library to select for novel antibodies against four human protein targets and demonstrate that identification of enriched sequences at each of the six CDRs in early selection rounds can be used to reconstruct a consensus antibody with selectivity for the target.
authors.affiliation: AxioMx, Inc., 688 East Main Street, Branford, CT 06405, United States. Electronic address: mbatonick@axiomxinc.com.
authors.forename: Melissa
authors.initials: M
authors.lastname: Batonick
authors.affiliation: AxioMx, Inc., 688 East Main Street, Branford, CT 06405, United States.
authors.forename: Erika G
authors.initials: EG
authors.lastname: Holland
authors.affiliation: AxioMx, Inc., 688 East Main Street, Branford, CT 06405, United States.
authors.forename: Valeria
authors.initials: V
authors.lastname: Busygina
authors.affiliation: AxioMx, Inc., 688 East Main Street, Branford, CT 06405, United States.
authors.forename: Dawn
authors.initials: D
authors.lastname: Alderman
authors.affiliation: University of Illinois at Chicago, 845 West Taylor Street Chicago, IL 60607, United States.
authors.forename: Brian K
authors.initials: BK
authors.lastname: Kay
authors.affiliation: AxioMx, Inc., 688 East Main Street, Branford, CT 06405, United States.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Weiner
authors.affiliation: AxioMx, Inc., 688 East Main Street, Branford, CT 06405, United States.
authors.forename: Margaret M
authors.initials: MM
authors.lastname: Kiss
mesh_terms: D000595:Amino Acid Sequence; D000916:Antibody Diversity; D001709:Biotechnology; D022801:Complementarity Determining Regions / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D057166:High-Throughput Screening Assays; D006801:Humans; D019151:Peptide Library; D057127:Single-Chain Antibodies / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D022801:Complementarity Determining Regions; D019151:Peptide Library; D057127:Single-Chain Antibodies