Development and differentiation of hyperstable consensus monobodies based on the type 3 fibronectin domain

Monobodies based on the type 3 fibronectin (FN3) domain are designed to overcome limitations of the antibody binding scaffold, being more simple and more amenable to the rigours of drug development. This thesis focuses on the development of FN3Con, a hyperstable consensus FN3 monobody. Through grafting of binding loops from monobodies of median stability, this work produced two FN3Con constructs with affinity to clinical targets and a shelf life of over 2 years at room temperature. The thesis then investigates methods to meaningfully differentiate monobodies from the antibody scaffold through buffer excipients, chemical conjugation and integration of native fibronectin characteristics.