title: Discovery of high affinity anti-ricin antibodies by B cell receptor sequencing and by yeast display of combinatorial VH:VL libraries from immunized animals.
abstract: Ricin is a toxin that could potentially be used as a bioweapon. We identified anti-ricin A chain antibodies by sequencing the antibody repertoire from immunized mice and by selecting high affinity antibodies using yeast surface display. These methods led to the isolation of multiple antibodies with high (sub-nanomolar) affinity. Interestingly, the antibodies identified by the 2 independent approaches are from the same clonal lineages, indicating for the first time that yeast surface display can identify native antibodies. The new antibodies represent well-characterized reagents for biodefense diagnostics and therapeutics development.
authors.affiliation: a Department of Chemical Engineering , University of Texas at Austin , Austin , TX , USA.
authors.forename: Bo
authors.initials: B
authors.lastname: Wang
authors.affiliation: a Department of Chemical Engineering , University of Texas at Austin , Austin , TX , USA.
authors.forename: Chang-Han
authors.initials: CH
authors.lastname: Lee
authors.affiliation: a Department of Chemical Engineering , University of Texas at Austin , Austin , TX , USA.
authors.forename: Erik L
authors.initials: EL
authors.lastname: Johnson
authors.affiliation: b Department of Molecular Biosciences , University of Texas at Austin , Austin , TX , USA.
authors.forename: Christien A
authors.initials: CA
authors.lastname: Kluwe
authors.affiliation: b Department of Molecular Biosciences , University of Texas at Austin , Austin , TX , USA.
authors.forename: Josephine C
authors.initials: JC
authors.lastname: Cunningham
authors.affiliation: a Department of Chemical Engineering , University of Texas at Austin , Austin , TX , USA.
authors.forename: Hidetaka
authors.initials: H
authors.lastname: Tanno
authors.affiliation: b Department of Molecular Biosciences , University of Texas at Austin , Austin , TX , USA.
authors.forename: Richard M
authors.initials: RM
authors.lastname: Crooks
authors.affiliation: a Department of Chemical Engineering , University of Texas at Austin , Austin , TX , USA.|b Department of Molecular Biosciences , University of Texas at Austin , Austin , TX , USA.|c Center for Systems and Synthetic Biology , University of Texas at Austin , Austin , TX , USA.|d Institute for Cellular and Molecular Biology , University of Texas at Austin , Austin , TX , USA.|e Department of Biomedical Engineering , University of Texas at Austin , Austin , TX , USA.
authors.forename: George
authors.initials: G
authors.lastname: Georgiou
authors.affiliation: b Department of Molecular Biosciences , University of Texas at Austin , Austin , TX , USA.|c Center for Systems and Synthetic Biology , University of Texas at Austin , Austin , TX , USA.|d Institute for Cellular and Molecular Biology , University of Texas at Austin , Austin , TX , USA.
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Ellington
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D000915:Antibody Affinity; D001402:B-Lymphocytes / Q000276:immunology; D001854:Bone Marrow Cells / Q000166:cytology; D061505:Cell Surface Display Techniques; D005260:Female; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D007114:Immunization; D008198:Lymph Nodes / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D019151:Peptide Library; D012333:RNA, Messenger / Q000235:genetics; D011947:Receptors, Antigen, B-Cell / Q000737:chemistry / Q000235:genetics*; D012276:Ricin / Q000276:immunology*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D061905:Single-Domain Antibodies / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D013154:Spleen / Q000166:cytology
chemical_list: D000906:Antibodies; D019151:Peptide Library; D012333:RNA, Messenger; D011947:Receptors, Antigen, B-Cell; D061905:Single-Domain Antibodies; D012276:Ricin
keywords: Antibody repertoire analysis; next-generation sequencing; ricin A chain; yeast surface display