Generation of high-affinity human antibodies by combining donor-derived and synthetic complementarity-determining-region diversity


Combinatorial libraries of rearranged hypervariable VH and VL sequences from nonimmunized human donors contain antigen specificities, including anti-self reactivities, created by random pairing of VHs and VLs. 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 libraries1,2. 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 antibodies3,4,5,6. This very often circumvents the need for affinity maturation, accelerating discovery of antibody drug candidates.

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Figure 1: Design of Dyax libraries.
Figure 2: Fab dissociation constants (Kd) in nM for selected Fabs.
Figure 3: Diversity observed in the new libraries compared to diversity seen in the PCR library.

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We are very grateful to Ed Madison and colleagues at Dendreon for the provision of the DESC1 and MSPL antigens. In addition, we are indebted to Kari Alitalo for our collaboration on TIE-1. We thank all our colleagues at Dyax, in both Cambridge and Liege, for many contributions and discussions throughout the course of this work.

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Corresponding author

Correspondence to Robert Charles Ladner.

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Competing interests

All of the authors worked for Dyax Corp. during the work on the library and were paid by Dyax. Dyax licenses the library for financial consideration.

Supplementary information

Supplementary Fig. 1

Schematic representation of the novel ONCL technology used to clone the CDR3-heavy chain repertoire from autoimmune donors. (PDF 339 kb)

Supplementary Table 1

Human Fabs that bind human protein targets (PDF 375 kb)

Supplementary Table 2

t/c (XLS 25 kb)

Supplementary Table 3

Oligonucleotides (PDF 117 kb)

Supplementary Discussion (PDF 124 kb)

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Hoet, R., Cohen, E., Kent, R. et al. Generation of high-affinity human antibodies by combining donor-derived and synthetic complementarity-determining-region diversity. Nat Biotechnol 23, 344–348 (2005).

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