title: In vitro nanobody discovery for integral membrane protein targets.
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.
authors.affiliation: Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093.
authors.forename: Rupak
authors.initials: R
authors.lastname: Doshi
authors.affiliation: Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093.
authors.forename: Beverly R
authors.initials: BR
authors.lastname: Chen
authors.affiliation: Vibat Consulting, P.O. Box 2835, Del Mar, CA 92014.
authors.forename: Cecile Rose T
authors.initials: CR
authors.lastname: Vibat
authors.affiliation: Department of Bioengineering, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093.
authors.forename: Norman
authors.initials: N
authors.lastname: Huang
authors.affiliation: Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093.
authors.forename: Chang-Wook
authors.initials: CW
authors.lastname: Lee
authors.affiliation: 1] Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093 [2] Department of Pharmacology, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093.
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Chang
mesh_terms: D000915:Antibody Affinity / Q000276:immunology; D061505:Cell Surface Display Techniques; D015870:Gene Expression; D015723:Gene Library; D017930:Genes, Reporter; D051272:Glucose Transporter Type 1 / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011485:Protein Binding; D012333:RNA, Messenger / Q000235:genetics; D011993:Recombinant Fusion Proteins; D061905:Single-Domain Antibodies / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D051272:Glucose Transporter Type 1; D008565:Membrane Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D061905:Single-Domain Antibodies