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
based recombinant systems, and decreased aggregation propensity. The new
beta super-binders behaved as potent super agonists, both in vitro and in
vivo. The latter scenario showed their better preformance when compared to
both non-mutated IL-2 and H9. The current dataset contains source data for
graphics showing frequency mutations and charge distribution among
unselected variants contained in phage-displayed libraries and selected
clones enriched after selection on immobilized beta chain. Data showing
the direct comparison between different IL-2 mutated variants produced as
Fc-fusion proteins are also presented. The comparison includes the results
of beta chain binding assays (ELISA), proliferation and phosphorylation
assays in vitro, in vivo expansion of lymphocyte populations and
anti-tumor activity in animal models. Taken together, the above described
data support the unique features of the new beta super-binders and their
potential as immunostimulatory and anti-cancer agents.