Abstract
Nanobodies (Nbs) or single-domain antibodies are among the smallest and most stable binder scaffolds known. In vitro display is a powerful antibody discovery technique used worldwide. We describe the first adaptation of in vitro mRNA/cDNA display for the rapid, automatable discovery of Nbs against desired targets, and use it to discover the first ever reported nanobody against the human full-length glucose transporter, GLUT-1. We envision our streamlined method as a bench-top platform technology, in combination with various molecular evolution techniques, for expedited Nb discovery.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Antibody Affinity / immunology
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Cell Surface Display Techniques
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Gene Expression
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Gene Library
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Genes, Reporter
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Glucose Transporter Type 1 / genetics
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Glucose Transporter Type 1 / immunology
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Glucose Transporter Type 1 / isolation & purification
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Glucose Transporter Type 1 / metabolism
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Humans
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In Vitro Techniques
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Membrane Proteins / genetics
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Membrane Proteins / immunology*
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Membrane Proteins / metabolism
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Protein Binding
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RNA, Messenger / genetics
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Recombinant Fusion Proteins
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Single-Domain Antibodies / genetics
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Single-Domain Antibodies / immunology*
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Single-Domain Antibodies / metabolism
Substances
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Glucose Transporter Type 1
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Membrane Proteins
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RNA, Messenger
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Recombinant Fusion Proteins
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Single-Domain Antibodies