title: Diversity of the antibody response to tetanus toxoid: comparison of hybridoma library to phage display library.
abstract: Monoclonal antibodies are important tools in research and since the 1990s have been an important therapeutic class targeting a wide variety of diseases. Earlier methods of mAb production relied exclusively on the lengthy process of making hybridomas. The advent of phage display technology introduced an alternative approach for mAb production. A potential concern with this approach is its complete dependence on an in vitro selection process, which may result in selection of V(H)-V(L) pairs normally eliminated during the in vivo selection process. The diversity of V(H)-V(L) pairs selected from phage display libraries relative to an endogenous response is unknown. To address these questions, we constructed a panel of hybridomas and a phage display library using the spleen of a single tetanus toxoid-immunized mouse and compared the diversity of the immune response generated using each technique. Surprisingly, the tetanus toxoid-specific antibodies produced by the hybridoma library exhibited a higher degree of V(H)-V(L) genetic diversity than their phage display-derived counterparts. Furthermore, the overlap among the V-genes from each library was very limited. Consistent with the notion that accumulation of many small DNA changes lead to increased antigen specificity and affinity, the phage clones displayed substantial micro-heterogeneity. Contrary to previous reports, we found that antigen specificity against tetanus toxoid is encoded by both V(κ) and V(H) genes. Finally, the phage-derived tetanus-specific clones had a lower binding affinity than the hybridomas, a phenomenon thought to be the result of random pairing of the V-genes.
authors.affiliation: Laboratory of Molecular and Developmental Immunology, Division of Monoclonal Antibodies, Center for Drug Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, United States of America.
authors.forename: Mahsa
authors.initials: M
authors.lastname: Sorouri
authors.affiliation: Laboratory of Molecular and Developmental Immunology, Division of Monoclonal Antibodies, Center for Drug Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, United States of America.
authors.forename: Sean P
authors.initials: SP
authors.lastname: Fitzsimmons
authors.affiliation: Laboratory of Molecular and Developmental Immunology, Division of Monoclonal Antibodies, Center for Drug Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, United States of America.
authors.forename: Antonina G
authors.initials: AG
authors.lastname: Aydanian
authors.affiliation: Laboratory of Molecular and Developmental Immunology, Division of Monoclonal Antibodies, Center for Drug Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, United States of America.
authors.forename: Sonita
authors.initials: S
authors.lastname: Bennett
authors.affiliation: Laboratory of Molecular and Developmental Immunology, Division of Monoclonal Antibodies, Center for Drug Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, United States of America.
authors.forename: Marjorie A
authors.initials: MA
authors.lastname: Shapiro
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000915:Antibody Affinity; D000917:Antibody Formation; D000918:Antibody Specificity; D001483:Base Sequence; D061505:Cell Surface Display Techniques; D004926:Escherichia coli; D005260:Female; D006825:Hybridomas; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D013744:Tetanus Toxin / Q000276:immunology*; D050441:VDJ Exons
chemical_list: D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D013744:Tetanus Toxin