Abstract
Multi-pass membrane proteins are important targets of biologic medicines. Given the inherent difficulties in working with membrane proteins, we sought to investigate the utility of membrane scaffold protein nanodiscs as a means of solubilizing membrane proteins to aid antibody discovery. Using a model multi-pass membrane protein, we demonstrate how incorporation of a multi-pass membrane protein into nanodiscs can be used in flow cytometry to identify antigen-specific hybridoma. The use of target protein-loaded nanodiscs to sort individual hybridoma early in the screening process can reduce the time required to identify antibodies against multi-pass membrane proteins.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antibodies, Monoclonal / immunology*
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Antibody Specificity
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Antigen-Antibody Reactions
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Arcobacter / chemistry
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Bacterial Proteins / chemistry
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Drug Delivery Systems
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Flow Cytometry / methods*
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Hybridomas / cytology*
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Hybridomas / immunology
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Immunoglobulin G / immunology*
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Membrane Proteins / immunology*
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Mice
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Models, Molecular
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NAV1.7 Voltage-Gated Sodium Channel / chemistry
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Nanostructures*
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Protein Conformation
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Protein Domains
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Recombinant Fusion Proteins / chemistry
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Solubility
Substances
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Antibodies, Monoclonal
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Bacterial Proteins
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Immunoglobulin G
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Membrane Proteins
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NAV1.7 Voltage-Gated Sodium Channel
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Recombinant Fusion Proteins
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SCN9A protein, human