title: Internal 32P-labeling of L-deoxyoligonucleotides.
abstract: A general two step procedure for the internal labeling of L-deoxyoligonucleotides, Spiegelmers, has been developed. Through radioactive labeling oligonucleotides can easily be detected and monitored in biological samples. T4 polynucleotide kinase is shown to efficiently phosphorylate strands of L-nucleic acids which allows the labeling with phosphorous isotopes such as (32)P. In order to protect the terminal phosphate label against unspecific phosphatases, one of two fragments of a Spiegelmer is enzymatically phosphorylated with [gamma-(32)P]ATP. In a second step we used a template- directed chemical ligation reaction in order to attach the labeled oligonucleotide to the other fragment to yield the full-length Spiegelmer with an internal [(32)P]phosphodiester bond. It has been shown that the functionality of a chemically ligated Spiegelmer is still retained.
authors.affiliation: NOXXON Pharma AG, Max-Dohrn-Strasse 8-10, D-10589 Berlin, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Frauendorf
authors.forename: Felix
authors.initials: F
authors.lastname: Hausch
authors.forename: Ingo
authors.initials: I
authors.lastname: Röhl
authors.forename: Andrea
authors.initials: A
authors.lastname: Lichte
authors.forename: Stefan
authors.initials: S
authors.lastname: Vonhoff
authors.forename: Sven
authors.initials: S
authors.lastname: Klussmann
mesh_terms: D017122:Bacteriophage T4 / Q000201:enzymology; D007553:Isotope Labeling / Q000379:methods*; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000378:metabolism*; D010761:Phosphorus Radioisotopes / Q000378:metabolism; D010766:Phosphorylation; D011116:Polynucleotide 5'-Hydroxyl-Kinase / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D009841:Oligonucleotides; D010761:Phosphorus Radioisotopes; D011116:Polynucleotide 5'-Hydroxyl-Kinase