A cell-based multiplex immunoassay platform using fluorescent protein-barcoded reporter cell lines.
| dc.contributor.author | Song, Shengli | |
| dc.contributor.author | Manook, Miriam | |
| dc.contributor.author | Kwun, Jean | |
| dc.contributor.author | Jackson, Annette M | |
| dc.contributor.author | Knechtle, Stuart J | |
| dc.contributor.author | Kelsoe, Garnett | |
| dc.date.accessioned | 2022-11-01T19:32:05Z | |
| dc.date.available | 2022-11-01T19:32:05Z | |
| dc.date.issued | 2021-11 | |
| dc.date.updated | 2022-11-01T19:31:41Z | |
| dc.description.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. | |
| dc.identifier | 10.1038/s42003-021-02881-w | |
| dc.identifier.issn | 2399-3642 | |
| dc.identifier.issn | 2399-3642 | |
| dc.identifier.uri | ||
| dc.language | eng | |
| dc.publisher | Springer Science and Business Media LLC | |
| dc.relation.ispartof | Communications biology | |
| dc.relation.isversionof | 10.1038/s42003-021-02881-w | |
| dc.subject | Antibodies | |
| dc.subject | Cell Line | |
| dc.subject | Epitopes | |
| dc.subject | Flow Cytometry | |
| dc.subject | Fluorescent Dyes | |
| dc.subject | Hemagglutinins | |
| dc.subject | Immunoassay | |
| dc.subject | Influenza A virus | |
| dc.subject | Membrane Proteins | |
| dc.subject | Mutation | |
| dc.subject | Protein Multimerization | |
| dc.subject | Receptors, CCR2 | |
| dc.subject | Receptors, CCR5 | |
| dc.title | A cell-based multiplex immunoassay platform using fluorescent protein-barcoded reporter cell lines. | |
| dc.type | Journal article | |
| duke.contributor.orcid | Kwun, Jean|0000-0002-8563-5472 | |
| duke.contributor.orcid | Jackson, Annette M|0000-0003-2648-2944 | |
| duke.contributor.orcid | Knechtle, Stuart J|0000-0002-1625-385X | |
| duke.contributor.orcid | Kelsoe, Garnett|0000-0002-8770-040X | |
| pubs.begin-page | 1338 | |
| pubs.issue | 1 | |
| pubs.organisational-group | Duke | |
| pubs.organisational-group | School of Medicine | |
| pubs.organisational-group | Basic Science Departments | |
| pubs.organisational-group | Clinical Science Departments | |
| pubs.organisational-group | Institutes and Centers | |
| pubs.organisational-group | Immunology | |
| pubs.organisational-group | Medicine | |
| pubs.organisational-group | Surgery | |
| pubs.organisational-group | Medicine, Nephrology | |
| pubs.organisational-group | Surgery, Abdominal Transplant Surgery | |
| pubs.organisational-group | Surgery, Surgical Sciences | |
| pubs.organisational-group | Duke Cancer Institute | |
| pubs.organisational-group | Duke Clinical Research Institute | |
| pubs.organisational-group | Duke Human Vaccine Institute | |
| pubs.publication-status | Published | |
| pubs.volume | 4 |
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