Generation of high-affinity human antibodies by combining donor-derived and synthetic complementarity-determining-region diversity.
Journal Article (Letter)
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.
Full Text
Duke Authors
Cited Authors
- Hoet, RM; Cohen, EH; Kent, RB; Rookey, K; Schoonbroodt, S; Hogan, S; Rem, L; Frans, N; Daukandt, M; Pieters, H; van Hegelsom, R; Neer, NC-V; Nastri, HG; Rondon, IJ; Leeds, JA; Hufton, SE; Huang, L; Kashin, I; Devlin, M; Kuang, G; Steukers, M; Viswanathan, M; Nixon, AE; Sexton, DJ; Hoogenboom, HR; Ladner, RC
Published Date
- March 2005
Published In
Volume / Issue
- 23 / 3
Start / End Page
- 344 - 348
PubMed ID
- 15723048
International Standard Serial Number (ISSN)
- 1087-0156
Digital Object Identifier (DOI)
- 10.1038/nbt1067
Language
- eng
Conference Location
- United States