A cell-based multiplex immunoassay platform using fluorescent protein-barcoded reporter cell lines.
Abstract
Multiplex immunoassays with acellular antigens are well-established based on solid-phase
platforms such as the Luminex® technology. Cell barcoding by amine-reactive fluorescent dyes enables analogous cell-based
multiplex assays, but requires multiple labeling reactions and quality checks prior
to every assay. Here we describe generation of stable, fluorescent protein-barcoded
reporter cell lines suitable for multiplex screening of antibody to membrane proteins.
The utility of this cell-based system, with the potential of a 256-plex cell panel,
is demonstrated by flow cytometry deconvolution of barcoded cell panels expressing
influenza A hemagglutinin trimers, or native human CCR2 or CCR5 multi-span proteins
and their epitope-defining mutants. This platform will prove useful for characterizing
immunity and discovering antibodies to membrane-associated proteins.
Type
Journal articleSubject
AntibodiesCell Line
Epitopes
Flow Cytometry
Fluorescent Dyes
Hemagglutinins
Immunoassay
Influenza A virus
Membrane Proteins
Mutation
Protein Multimerization
Receptors, CCR2
Receptors, CCR5
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https://hdl.handle.net/10161/26168Published Version (Please cite this version)
10.1038/s42003-021-02881-wPublication Info
Song, Shengli; Manook, Miriam; Kwun, Jean; Jackson, Annette M; Knechtle, Stuart J;
& Kelsoe, Garnett (2021). A cell-based multiplex immunoassay platform using fluorescent protein-barcoded reporter
cell lines. Communications biology, 4(1). pp. 1338. 10.1038/s42003-021-02881-w. Retrieved from https://hdl.handle.net/10161/26168.This is constructed from limited available data and may be imprecise. To cite this
article, please review & use the official citation provided by the journal.
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Show full item recordScholars@Duke
Annette M Jackson
Associate Professor in Surgery
Garnett H. Kelsoe
James B. Duke Distinguished Professor of Immunology
1. Lymphocyte development and antigen-driven diversification of immunoglobulin and
T cell antigen receptor genes. 2. The germinal center reaction and mechanisms for
clonal selection and self - tolerance. The origins of autoimmunity. 3. Interaction
of innate- and adaptive immunity and the role of inflammation in lymphoid organogenesis.
4. The role of secondary V(D)J gene rearrangment in lymphocyte development and malignancies.
5. Mathematical modeling of immune responses,
Stuart Johnston Knechtle
William R. Kenan, Jr. Distinguished Professor
During my career as an academic surgeon, I have had the privilege of leading and/or
participating in a diverse portfolio of hypothesis-driven research projects. These
projects have centered on the immunology of surgery and transplantation, including
both cellular and antibody-mediated immune responses. During my training I studied
the response of hyper-sensitized recipients to allogeneic liver transplantation, and
am currently studying means of reducing immunologic memory that might
Jean Kwun
Associate Professor in Surgery
Research interests include humoral tolerance to organ transplants in animal model
and humans, developing a clinically relevant animal model to study the mechanisms
of antibody-mediated rejection (AMR), and establishing a conceptual basis that will
translate into therapeutic intervention of AMR.
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