title: Magnetically labeled secretin retains receptor affinity to pancreas acinar cells.
abstract: Previously, we have developed a colloidal dextran-stabilized monocrystalline iron oxide nanocompound (MION-46) as a magnetic label for magnetic resonance imaging (MRI). In an effort to use this magnetic label to visualize pancreatic receptor function by MRI in vivo, we investigated the potential of secretin as a vector molecule. Secretin receptors, abundant on exocrine pancreas cells, recognize secretin through its amidated carboxyl terminal. In order to conjugate secretin to MION, we utilized the specific interaction between biotin and streptavidin, since direct conjugation of human secretin to MION has previously resulted in low yields and low affinity of the conjugate (unpublished results). Initially, we biotinylated the N-terminal primary amino group of secretin (60% yield). In a separate step, streptavidin (SA) was immobilized onto the surface dextran molecules of MION (79% yield) by reductive amination. Each secretin molecule was conjugated to one biotin molecule and each MION particle to an average of two SA molecules. The biotinylated secretin was then conjugated to MION through the biotin-streptavidin interaction (90% yield). The secretin-biotin-streptavidin-MION construct thus contained approximately two secretin molecules per MION. An in vitro competitive binding assay of pancreatic acinar cells demonstrated that the magnetically labeled secretin retained affinity to the secretin receptors. In vivo distribution studies in rats showed a significantly higher pancreatic accumulation of the secretin-biotin-streptavidin-MION construct as compared to the control group that had received unmodified MION. Our data indicate that bioactive peptides can be attached to dextran-coated iron oxide particles through the biotin-streptavidin interaction while retaining receptor affinity. Such target-specific agents have potential use in MR imaging to probe for a variety of receptor systems.
authors.affiliation: Department of Radiology, Massachusetts General Hospital, Boston 02114, USA.
authors.forename: T T
authors.initials: TT
authors.lastname: Shen
authors.forename: A
authors.initials: A
authors.lastname: Bogdanov
authors.forename: A
authors.initials: A
authors.lastname: Bogdanova
authors.forename: K
authors.initials: K
authors.lastname: Poss
authors.forename: T J
authors.initials: TJ
authors.lastname: Brady
authors.forename: R
authors.initials: R
authors.lastname: Weissleder
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry; D001667:Binding, Competitive; D001710:Biotin / Q000737:chemistry; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003287:Contrast Media / Q000737:chemistry; D003911:Dextrans / Q000737:chemistry; D052203:Ferrosoferric Oxide; D006801:Humans; D007457:Iodine Radioisotopes; D007501:Iron / Q000032:analysis / Q000737:chemistry*; D007553:Isotope Labeling / Q000379:methods; D008279:Magnetic Resonance Imaging / Q000379:methods; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D010087:Oxides / Q000032:analysis / Q000737:chemistry*; D010179:Pancreas / Q000166:cytology / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D051381:Rats; D017207:Rats, Sprague-Dawley; D012633:Secretin / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D019809:Streptavidin; D014018:Tissue Distribution
chemical_list: D001426:Bacterial Proteins; D003287:Contrast Media; D003911:Dextrans; D007457:Iodine Radioisotopes; D010087:Oxides; D012633:Secretin; D001710:Biotin; D019809:Streptavidin; D007501:Iron; D052203:Ferrosoferric Oxide