title: Generation of high-affinity human antibodies by combining donor-derived and synthetic complementarity-determining-region diversity.
abstract: Combinatorial libraries of rearranged hypervariable V(H) and V(L) sequences from nonimmunized human donors contain antigen specificities, including anti-self reactivities, created by random pairing of V(H)s and V(L)s. Somatic hypermutation of immunoglobulin genes, however, is critical in the generation of high-affinity antibodies in vivo and occurs only after immunization. Thus, in combinatorial phage display libraries from nonimmunized donors, high-affinity antibodies are rarely found. Lengthy in vitro affinity maturation is often needed to improve antibodies from such libraries. We report the construction of human Fab libraries having a unique combination of immunoglobulin sequences captured from human donors and synthetic diversity in key antigen contact sites in heavy-chain complementarity-determining regions 1 and 2. The success of this strategy is demonstrated by identifying many monovalent Fabs against multiple therapeutic targets that show higher affinities than approved therapeutic antibodies. This very often circumvents the need for affinity maturation, accelerating discovery of antibody drug candidates.
authors.forename: René Michael
authors.initials: RM
authors.lastname: Hoet
authors.forename: Edward H
authors.initials: EH
authors.lastname: Cohen
authors.forename: Rachel Baribault
authors.initials: RB
authors.lastname: Kent
authors.forename: Kristin
authors.initials: K
authors.lastname: Rookey
authors.forename: Sonia
authors.initials: S
authors.lastname: Schoonbroodt
authors.forename: Shannon
authors.initials: S
authors.lastname: Hogan
authors.forename: Louise
authors.initials: L
authors.lastname: Rem
authors.forename: Nicolas
authors.initials: N
authors.lastname: Frans
authors.forename: Marc
authors.initials: M
authors.lastname: Daukandt
authors.forename: Henk
authors.initials: H
authors.lastname: Pieters
authors.forename: Rob
authors.initials: R
authors.lastname: van Hegelsom
authors.forename: Nicole Coolen-van
authors.initials: NC
authors.lastname: Neer
authors.forename: Horacio G
authors.initials: HG
authors.lastname: Nastri
authors.forename: Isaac J
authors.initials: IJ
authors.lastname: Rondon
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Leeds
authors.forename: Simon E
authors.initials: SE
authors.lastname: Hufton
authors.forename: Lili
authors.initials: L
authors.lastname: Huang
authors.forename: Irina
authors.initials: I
authors.lastname: Kashin
authors.forename: Mary
authors.initials: M
authors.lastname: Devlin
authors.forename: Guannan
authors.initials: G
authors.lastname: Kuang
authors.forename: Mieke
authors.initials: M
authors.lastname: Steukers
authors.forename: Malini
authors.initials: M
authors.lastname: Viswanathan
authors.forename: Andrew E
authors.initials: AE
authors.lastname: Nixon
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Sexton
authors.forename: Hennie R
authors.initials: HR
authors.lastname: Hoogenboom
authors.forename: Robert Charles
authors.initials: RC
authors.lastname: Ladner
mesh_terms: D000915:Antibody Affinity; D000917:Antibody Formation; D022801:Complementarity Determining Regions / Q000235:genetics*; D014644:Genetic Variation / Q000235:genetics; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D019151:Peptide Library; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods*; D011995:Recombination, Genetic / Q000235:genetics; D014019:Tissue Donors
chemical_list: D022801:Complementarity Determining Regions; D007140:Immunoglobulin Fab Fragments; D019151:Peptide Library