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ACS Nano. 2016 Dec 27;10(12):10652-10660. doi: 10.1021/acsnano.6b03786. Epub 2016 Sep 20.

High-Resolution Analysis of Antibodies to Post-Translational Modifications Using Peptide Nanosensor Microarrays.

Author information

1
Intel Corporation , Santa Clara, California 95052, United States.
2
Department of Molecular and Cell Biology, Division of Immunology and Pathogenesis, University of California , Berkeley, California 94720, United States.
3
Department of Electrical and Computer Engineering, University of California , San Diego, California 92093, United States.
4
Center for Immunology, University of Minnesota Medical School , Minneapolis, Minnesota 55455, United States.
5
Division of Rheumatology, Johns Hopkins University School of Medicine , Baltimore, Maryland 21205, United States.
6
Institute for Immunity, Transplantation, and Infection, Stanford University School of Medicine , Stanford, California 94305, United States.

Abstract

Autoantibodies are a hallmark of autoimmune diseases such as lupus and have the potential to be used as biomarkers for diverse diseases, including immunodeficiency, infectious disease, and cancer. More precise detection of antibodies to specific targets is needed to improve diagnosis of such diseases. Here, we report the development of reusable peptide microarrays, based on giant magnetoresistive (GMR) nanosensors optimized for sensitively detecting magnetic nanoparticle labels, for the detection of antibodies with a resolution of a single post-translationally modified amino acid. We have also developed a chemical regeneration scheme to perform multiplex assays with a high level of reproducibility, resulting in greatly reduced experimental costs. In addition, we show that peptides synthesized directly on the nanosensors are approximately two times more sensitive than directly spotted peptides. Reusable peptide nanosensor microarrays enable precise detection of autoantibodies with high resolution and sensitivity and show promise for investigating antibody-mediated immune responses to autoantigens, vaccines, and pathogen-derived antigens as well as other fundamental peptide-protein interactions.

KEYWORDS:

autoantibody; giant magnetoresistance; lupus; nanosensors; peptide microarray; regeneration

PMID:
27636738
PMCID:
PMC5367622
DOI:
10.1021/acsnano.6b03786
[Indexed for MEDLINE]
Free PMC Article

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