title: Assessing kinetic and epitopic diversity across orthogonal monoclonal antibody generation platforms.
abstract: The ability of monoclonal antibodies (mAbs) to target specific antigens with high precision has led to an increasing demand to generate them for therapeutic use in many disease areas. Historically, the discovery of therapeutic mAbs has relied upon the immunization of mammals and various in vitro display technologies. While the routine immunization of rodents yields clones that are stable in serum and have been selected against vast arrays of endogenous, non-target self-antigens, it is often difficult to obtain species cross-reactive mAbs owing to the generally high sequence similarity shared across human antigens and their mammalian orthologs. In vitro display technologies bypass this limitation, but lack an in vivo screening mechanism, and thus may potentially generate mAbs with undesirable binding specificity and stability issues. Chicken immunization is emerging as an attractive mAb discovery method because it combines the benefits of both in vivo and in vitro display methods. Since chickens are phylogenetically separated from mammals, their proteins share less sequence homology with those of humans, so human proteins are often immunogenic and can readily elicit rodent cross-reactive clones, which are necessary for in vivo proof of mechanism studies. Here, we compare the binding characteristics of mAbs isolated from chicken immunization, mouse immunization, and phage display of human antibody libraries. Our results show that chicken-derived mAbs not only recapitulate the kinetic diversity of mAbs sourced from other methods, but appear to offer an expanded repertoire of epitopes. Further, chicken-derived mAbs can bind their native serum antigen with very high affinity, highlighting their therapeutic potential.
authors.affiliation: a Protein Engineering, Rinat-Pfizer Inc. , South San Francisco , CA , USA.
authors.forename: Yasmina Noubia
authors.initials: YN
authors.lastname: Abdiche
authors.affiliation: b Crystal Bioscience , CA , USA.
authors.forename: Rian
authors.initials: R
authors.lastname: Harriman
authors.affiliation: a Protein Engineering, Rinat-Pfizer Inc. , South San Francisco , CA , USA.
authors.forename: Xiaodi
authors.initials: X
authors.lastname: Deng
authors.affiliation: a Protein Engineering, Rinat-Pfizer Inc. , South San Francisco , CA , USA.
authors.forename: Yik Andy
authors.initials: YA
authors.lastname: Yeung
authors.affiliation: c Wasatch Microfluidics , Salt Lake City , UT , USA.
authors.forename: Adam
authors.initials: A
authors.lastname: Miles
authors.affiliation: a Protein Engineering, Rinat-Pfizer Inc. , South San Francisco , CA , USA.
authors.forename: Winse
authors.initials: W
authors.lastname: Morishige
authors.affiliation: a Protein Engineering, Rinat-Pfizer Inc. , South San Francisco , CA , USA.
authors.forename: Leila
authors.initials: L
authors.lastname: Boustany
authors.affiliation: b Crystal Bioscience , CA , USA.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhu
authors.affiliation: b Crystal Bioscience , CA , USA.
authors.forename: Shelley Mettler
authors.initials: SM
authors.lastname: Izquierdo
authors.affiliation: b Crystal Bioscience , CA , USA.
authors.forename: William
authors.initials: W
authors.lastname: Harriman
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D030161:Avian Proteins / Q000276:immunology*; D001666:Binding Sites, Antibody; D002645:Chickens / Q000276:immunology*; D000939:Epitopes / Q000276:immunology*; D005260:Female; D006801:Humans; D007700:Kinetics; D051379:Mice; D013045:Species Specificity
chemical_list: D000911:Antibodies, Monoclonal; D030161:Avian Proteins; D000939:Epitopes
keywords: Binning; Immunology; chicken immune repertoire; epitope; monoclonal antibody