Molecular reshaping of phage-displayed Interleukin-2 at beta chain receptor interface to obtain potent super-agonists with improved developability profiles-primary dataset

 

Interleukin-2 (IL-2) had been been engineered up to now by yeast display and in silico rational design. In this article we reshaped IL-2 interface with the IL-2 receptor beta subunit to increase binding affinity between both interacting partners, using phage display. Multiple IL-2 mutated variants were selected from large phage-displayed libraries, showing shared molecular patterns. An accumulation of negative charges in the segment 81-87 of IL-2 primary sequence was observed, as well as the strong preponderance of the replacement I92L. The first feature contributed to an optimized electrostatic complementarity between IL-2 and IL-2 receptor beta chain, resulting in higher affinity and faster association kinetics than the ones of previously reported H9 superkine retrieved from yeast display libraries. The presence of a Leu residue at position 92 was the key molecular determinant for a favourable biophysical profile characterized by high stability and production in mammalian-cell...