Generation of high-affinity human antibodies by combining donor-derived and synthetic complementarity-determining-region diversity.
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.
Hoet
René Michael
RM
Cohen
Edward H
EH
Kent
Rachel Baribault
RB
Rookey
Kristin
K
Schoonbroodt
Sonia
S
Hogan
Shannon
S
Rem
Louise
L
Frans
Nicolas
N
Daukandt
Marc
M
Pieters
Henk
H
van Hegelsom
Rob
R
Neer
Nicole Coolen-van
NC
Nastri
Horacio G
HG
Rondon
Isaac J
IJ
Leeds
Jennifer A
JA
Hufton
Simon E
SE
Huang
Lili
L
Kashin
Irina
I
Devlin
Mary
M
Kuang
Guannan
G
Steukers
Mieke
M
Viswanathan
Malini
M
Nixon
Andrew E
AE
Sexton
Daniel J
DJ
Hoogenboom
Hennie R
HR
Ladner
Robert Charles
RC
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
D022801:Complementarity Determining Regions; D007140:Immunoglobulin Fab Fragments; D019151:Peptide Library