title: Transfected cell microarrays for the expression of membrane-displayed single-chain antibodies.
abstract: Transfected cell microarrays, arrays of cells expressing defined cDNAs, are promising technologies that can enable the functional analysis of many proteins in parallel. This technique has been adapted for the comparative functional analysis of single-chain antibodies (scFvs) and to facilitate the screening and characterization of these antibodies for their use in diagnostic and therapeutic applications. In this method, membrane-targeting expression vectors encoding scFvs are mixed with transfection reagents and are deposited at high density onto a microscope slide. Adherent mammalian cells are subsequently added to the printed array. Upon attachment to the substrate, the cells take up the plasmid DNA and express the particular protein encoded at each location. The result is an array whose features are micrometer-sized clusters of cells expressing defined genes. This approach provides for the high-throughput functional analysis of many different proteins in parallel and can be considerably more informative and cost-effective relative to more traditional protein expression techniques.
authors.affiliation: United States Naval Research Laboratory, Center for Bio/Molecular Science and Engineering, Washington, DC, USA. baochuan.lin@nrl.navy.mil
authors.forename: Baochuan
authors.initials: B
authors.lastname: Lin
authors.forename: James B
authors.initials: JB
authors.lastname: Delehanty
mesh_terms: D000906:Antibodies / Q000235:genetics; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D005822:Genetic Vectors; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D007211:Indoles / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D046888:Tissue Array Analysis / Q000379:methods*; D014162:Transfection
chemical_list: D000906:Antibodies; D005456:Fluorescent Dyes; D007211:Indoles; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies; C007293:DAPI; D004247:DNA; D016650:Fluorescein-5-isothiocyanate