Miller, Eric A. Sung, Ki-Joo Kongsuphol, Patthara Baniya, Subha Aw-Yong, Hui Qi Tay, Vivian Tan, Yuxuan Kabir, Farah M. Pang-Yeo, Karl Kaspriskie, Isabel G. Sikes, Hadley D. Beyond Epitope Binning: Directed <i>in Vitro</i> Selection of Complementary Pairs of Binding Proteins Many 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 <i>post facto</i> 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 <i>in vitro</i> libraries of ∼10<sup>9</sup> 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. affinity pairs;RAPIDS;nonoverlapping target epitopes;Rapid Affinity Pair Identification;affinity reagents 2019-12-24
    https://acs.figshare.com/articles/journal_contribution/Beyond_Epitope_Binning_Directed_i_in_Vitro_i_Selection_of_Complementary_Pairs_of_Binding_Proteins/11449200
10.1021/acscombsci.9b00176.s001