title: Dual isotope labeling: conjugation of 32P-oligonucleotides with 18F-aryltrifluoroborate via copper(I) catalyzed cycloaddition.
abstract: A one-pot-two-step labeling of an oligonucleotide with an (18)F-ArBF3(-)(aryltrifluoroborate) radioprosthetic is reported herein. In order to characterize labeling in terms of radiochemistry, phosphorus-32 was also introduced to the 5'-terminus of the oligonucleotide via enzymatic phosphorylation. A pendant azide group was subsequently conjugated to the 5'-phosphate of the oligonucleotide. Copper(I) catalyzed [2+3] cycloaddition was undertaken to conjugate an alkyne-bearing(18)F-ArBF3(-) to the oligonucleotide. Following polyacrylamide gel electrophoresis, this doubly-labeled bioconjugate exhibited decay properties of both the phosphorus-32 and fluorine-18, that were confirmed by autoradiography at selected lengths of time, which in turn provided concrete evidence of successful conjugation. These results are corroborated by HPLC analysis of the labeled material. Taken together this work demonstrates viable use of (18)F-ArBF3(-) prosthetics for labeling oligonucleotides for use in PET imaging.
authors.affiliation: Jiangsu Key Laboratory of Atmospheric Environment Monitoring & Pollution Control, College of Environmental Science & Engineering, Nanjing University of Information Science & Technology, 219 Ningliu Road, Nanjing 210044, PR China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Li
authors.forename: Paul
authors.initials: P
authors.lastname: Schaffer
authors.forename: David M
authors.initials: DM
authors.lastname: Perrin
mesh_terms: D000480:Alkynes / Q000737:chemistry; D001881:Borates / Q000138:chemical synthesis* / Q000737:chemistry; D002414:Cations, Monovalent / Q000737:chemistry; D057930:Click Chemistry; D003300:Copper / Q000737:chemistry*; D061565:Cycloaddition Reaction; D005462:Fluorine Radioisotopes / Q000737:chemistry*; D007553:Isotope Labeling / Q000379:methods; D015394:Molecular Structure; D009841:Oligonucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D010761:Phosphorus Radioisotopes / Q000737:chemistry*; D013237:Stereoisomerism
chemical_list: D000480:Alkynes; D001881:Borates; D002414:Cations, Monovalent; D005462:Fluorine Radioisotopes; D009841:Oligonucleotides; D010761:Phosphorus Radioisotopes; D003300:Copper
keywords: Aryltrifluoroborates (ArBF(3)(−)s); Copper(I) catalyzed [2+3] cycloaddition; Fluorine-18; Oligonucleotides; Phosphorus-32