title: Cell surface expression of single chain antibodies with applications to imaging of gene expression in vivo.
abstract: Imaging of gene expression in vivo has many potential uses for biomedical research and drug discovery, ranging from the study of gene regulation and cancer to the non-invasive assessment of gene therapies. To streamline the development of imaging marker gene technologies for nuclear medicine, we propose a new approach to the design of reporter/probe pairs wherein the reporter is a cell surface-expressed single chain antibody variable fragment that has been raised against a low molecular weight imaging probe with optimized pharmacokinetic properties. Proof of concept of the approach was achieved using a single chain antibody variable fragment that binds with high affinity to fluorescein and an imaging probe consisting of fluorescein isothiocyanate coupled to the chelator diethylene triamine penta-acetic acid labeled with the gamma-emitter (111)In. We demonstrate specific high-affinity binding of this probe to the cell surface-expressed reporter in vitro and assess the in vivo biodistribution of the probe both in wild-type mice and in mice harboring tumor xenografts expressing the reporter. Specific uptake of the probe by, and in vivo imaging of, tumors expressing the reporter are shown. Since ScFvs with high affinities can be raised to almost any protein or small molecule, the proposed methodology may offer a new flexibility in the design of imaging tracer/reporter pairs wherein both probe pharmacokinetics and binding affinities can be readily optimized.
authors.affiliation: Affymax Research Institute, Palo Alto, CA, USA. Jeff.Northrop@PETNetPharmaceutical.com
authors.forename: Jeffrey P
authors.initials: JP
authors.lastname: Northrop
authors.forename: Mark
authors.initials: M
authors.lastname: Bednarski
authors.forename: Susan O
authors.initials: SO
authors.lastname: Barbieri
authors.forename: Amy T
authors.initials: AT
authors.lastname: Lu
authors.forename: Dee
authors.initials: D
authors.lastname: Nguyen
authors.forename: John
authors.initials: J
authors.lastname: Varadarajan
authors.forename: Maureen
authors.initials: M
authors.lastname: Osen
authors.forename: King C
authors.initials: KC
authors.lastname: Li
authors.forename: Josh
authors.initials: J
authors.lastname: Star-Lack
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008819:Mice, Nude; D011863:Radioimmunoassay / Q000379:methods*; D016719:Radioimmunodetection / Q000379:methods*; D019275:Radiopharmaceuticals / Q000276:immunology / Q000493:pharmacokinetics
chemical_list: D000906:Antibodies; D007128:Immunoglobulin Fragments; D019275:Radiopharmaceuticals; C055966:immunoglobulin Fv