Cluster of Excellence –
University of Freiburg

A two-step approach for the design and generation of nanobodies

Wagner HJ, Wehrle S, Weiss E, Cavallari M, Weber W.

Int J Mol Sci. 2018 Nov 2;19(11).

Int J Mol Sci.        online article

Here, we propose a two-step approach for the design and generation of nanobodies. In a first step, complementarity-determining regions (CDRs) are grafted from conventional antibody formats onto nanobody frameworks, generating weak antigen binders. In a second step, the weak binders serve as templates to design focused synthetic phage libraries for affinity maturation. We validated this approach by grafting toxin- and hapten-specific CDRs onto frameworks derived from variable domains of camelid heavy-chain-only antibodies (VHH). We then affinity matured the hapten binder via panning of a synthetic phage library.