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{
    "pk": 4847,
    "title": "Determination of Immune Signal–Receptors, PD-1 and PD-L1, Interactions in Solution using qFRET Technology",
    "subtitle": null,
    "abstract": "Human PD-L1 (programmed cell death 1 - ligand 1) is a transmembrane protein that is highlyexpressed on the membrane of cancer cells. It binds to inhibitory receptor PD-1 (programmed celldeath protein - 1), which is expressed on the surface of cytotoxic T cells. The interaction betweenPD-L1 and PD-1 reduces the effect of anti-tumor immune response and the strategy of blocking theirinteraction has been used for anti-cancer drug manufacture. Past studies isolated the extracellulardomain of PD-L1 for characterization of the structure. This study aims to recover, isolate, and purifythe insoluble PD-L1 protein (external domain), and study its binding interaction with PD-1 forthe development of an in vitro quantitative FRET (qFRET) assay. To report PD-L1/PD-1 bindinginteraction, fluorescent donor and acceptor pairs, CyPet and YPet were bound to PD-L1 and PD-1proteins respectively and qFRET was applied to assess the interaction of the two proteins basedentirely on fluorescence.The results provide evidence of recovery of the purified and refolded CyPet-Ext.PDL1 and YPet-Ext.PD1 proteins from cell pellets, shown in the coomassie stained SDS-PAGE gel. This study alsoshows that the proteins were able to be recovered through affinity chromatography under denaturingconditions. The qFRET technique showed that the acceptor, YPet-Ext.PD1, is interacting with thedonor, CyPet-Ext.PDL1. This study provides a novel method for better understanding the bindingmechanism of PD-L1/PD-1 that can be applied to other cell-surface protein interactions, as well as tostipulate a platform for small molecule inhibitor related drug screenings and production.",
    "language": "en",
    "license": null,
    "keywords": [
        {
            "word": "PD-1"
        },
        {
            "word": "PD-L1"
        },
        {
            "word": "Expression"
        },
        {
            "word": "Immunotherapy"
        },
        {
            "word": "Antibody"
        },
        {
            "word": "FRET"
        }
    ],
    "section": "Articles",
    "is_remote": true,
    "remote_url": "https://escholarship.org/uc/item/2d09435x",
    "frozenauthors": [
        {
            "first_name": "Mary",
            "middle_name": "",
            "last_name": "Nwangwu",
            "name_suffix": "",
            "institution": "",
            "department": "None"
        },
        {
            "first_name": "Jiayu",
            "middle_name": "",
            "last_name": "Liao",
            "name_suffix": "",
            "institution": "",
            "department": "None"
        }
    ],
    "date_submitted": "2018-05-25T07:43:34+09:00",
    "date_accepted": "2018-05-25T07:43:34+09:00",
    "date_published": "2018-01-01T09:00:00+09:00",
    "render_galley": null,
    "galleys": [
        {
            "label": "",
            "type": "",
            "path": "https://journalpub.escholarship.org/ucr_undergrad_research_j/article/4847/galley/2742/download/"
        }
    ]
}