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Proceedings Paper

Peptide library synthesis on spectrally encoded beads for multiplexed protein/peptide bioassays
Author(s): Huy Q. Nguyen; Kara Brower; Björn Harink; Brian Baxter; Kurt S. Thorn; Polly M. Fordyce
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Paper Abstract

Protein-peptide interactions are essential for cellular responses. Despite their importance, these interactions remain largely uncharacterized due to experimental challenges associated with their measurement. Current techniques (e.g. surface plasmon resonance, fluorescence polarization, and isothermal calorimetry) either require large amounts of purified material or direct fluorescent labeling, making high-throughput measurements laborious and expensive. In this report, we present a new technology for measuring antibody-peptide interactions in vitro that leverages spectrally encoded beads for biological multiplexing. Specific peptide sequences are synthesized directly on encoded beads with a 1:1 relationship between peptide sequence and embedded code, thereby making it possible to track many peptide sequences throughout the course of an experiment within a single small volume. We demonstrate the potential of these bead-bound peptide libraries by: (1) creating a set of 46 peptides composed of 3 commonly used epitope tags (myc, FLAG, and HA) and single amino-acid scanning mutants; (2) incubating with a mixture of fluorescently-labeled antimyc, anti-FLAG, and anti-HA antibodies; and (3) imaging these bead-bound libraries to simultaneously identify the embedded spectral code (and thus the sequence of the associated peptide) and quantify the amount of each antibody bound. To our knowledge, these data demonstrate the first customized peptide library synthesized directly on spectrally encoded beads. While the implementation of the technology provided here is a high-affinity antibody/protein interaction with a small code space, we believe this platform can be broadly applicable to any range of peptide screening applications, with the capability to multiplex into libraries of hundreds to thousands of peptides in a single assay.

Paper Details

Date Published: 28 February 2017
PDF: 11 pages
Proc. SPIE 10061, Microfluidics, BioMEMS, and Medical Microsystems XV, 100610Z (28 February 2017); doi: 10.1117/12.2254908
Show Author Affiliations
Huy Q. Nguyen, Stanford Univ. (United States)
Univ. of California, San Francisco (United States)
Kara Brower, Stanford Univ. (United States)
Björn Harink, Standford Univ. (United States)
Univ. of California, San Francisco (United States)
Brian Baxter, Univ. of California, San Francisco (United States)
Kurt S. Thorn, Univ. of California, San Francisco (United States)
Polly M. Fordyce, Stanford Univ. (United States)


Published in SPIE Proceedings Vol. 10061:
Microfluidics, BioMEMS, and Medical Microsystems XV
Bonnie L. Gray; Holger Becker, Editor(s)

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