Beyond Epitope Binning: Directed in Vitro Selection of Complementary Pairs of Binding Proteins
Year: 2019
Type: article
Abstract: 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 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.
Source: ACS Combinatorial Science
Language: English
Subfield: Radiology, Nuclear Medicine and Imaging
Field: Medicine
Domain: Health Sciences
Open Access status: hybrid
Estimated APC paid: $5,000
Funders: National Science Foundation, Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, National Research Foundation, National Research Foundation Singapore, National Institutes of Health, Deshpande Center for Technological Innovation, Massachusetts Institute of Technology, Tata Center for Technology and Design, Massachusetts Institute of Technology, National Cancer Institute, National Institute of General Medical Sciences
OpenAlex ID: W2989927501