Beyond Epitope Binning: Directed in Vitro Selection of Complementary Pairs of Binding Proteins
Name
acscombsci.9b00176.pdf
Description
Published version
Size
3.5 MB
Format
Adobe PDF
Checksum (MD5)
b2b21615c5779c27760b76bb4fb48098
Author(s)
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
Date Issued
2020
Journal
ACS Combinatorial Science
Publisher
American Chemical Society (ACS)
Version
Final published version
Abstract
© 2019 American Chemical Society. 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 â¼109 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.
MIT Department
Massachusetts Institute of Technology. Department of Chemical Engineering
Singapore-MIT Alliance in Research and Technology (SMART)
Terms of Use
Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1021/ACSCOMBSCI.9B00176
