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Beyond Epitope Binning: Directed in Vitro Selection of Complementary Pairs of Binding Proteins
journal contribution
posted on 2019-12-24, 13:12 authored by Eric A. Miller, Ki-Joo Sung, Patthara Kongsuphol, Subha Baniya, Hui Qi Aw-Yong, Vivian Tay, Yuxuan Tan, Farah M. Kabir, Karl Pang-Yeo, Isabel G. Kaspriskie, Hadley D. SikesMany biotechnological applications require the simultaneous
binding
of affinity reagents to nonoverlapping target epitopes, the most prominent
example being sandwich immunoassays. Typically, affinity pairs are
identified via post facto functional analysis of
clones that were not selected for complementarity. Here, we developed
the Rapid Affinity Pair Identification via Directed Selection (RAPIDS)
process, which enables the efficient identification of affinity reagents
that function together as complementary pairs, from in vitro libraries of ∼109 variants. We used RAPIDS to
develop highly specific affinity pairs against biomarkers of tuberculosis,
Zika virus, and sepsis. Without additional trial-and-error screening,
these affinity pairs exhibited utility in multiple assay formats.
The RAPIDS process applies selective pressure to hundreds of thousands
of potential affinity pairs to efficiently identify complementary
pairs that bind to separate epitopes without binding to one another
or nontargets, yielding diagnostic assays that are sensitive and specific
by design.