title: Electrospray ionisation-cleavable tandem nucleic acid mass tag-peptide nucleic acid conjugates: synthesis and applications to quantitative genomic analysis using electrospray ionisation-MS/MS.
abstract: The synthesis and characterization of isotopomer tandem nucleic acid mass tag-peptide nucleic acid (TNT-PNA) conjugates is described along with their use as electrospray ionisation-cleavable (ESI-Cleavable) hybridization probes for the detection and quantification of target DNA sequences by electrospray ionisation tandem mass spectrometry (ESI-MS/MS). ESI-cleavable peptide TNT isotopomers were introduced into PNA oligonucleotide sequences in a total synthesis approach. These conjugates were evaluated as hybridization probes for the detection and quantification of immobilized synthetic target DNAs using ESI-MS/MS. In these experiments, the PNA portion of the conjugate acts as a hybridization probe, whereas the peptide TNT is released in a collision-based process during the ionization of the probe conjugate in the electrospray ion source. The cleaved TNT acts as a uniquely resolvable marker to identify and quantify a unique target DNA sequence. The method should be applicable to a wide variety of assays requiring highly multiplexed, quantitative DNA/RNA analysis, including gene expression monitoring, genetic profiling and the detection of pathogens.
authors.affiliation: Trillion Genomics Ltd, Babraham Research Campus, Babraham, Cambridge CB2 4AT, UK.
authors.forename: Andrew
authors.initials: A
authors.lastname: Thompson
authors.forename: Mark
authors.initials: M
authors.lastname: Prescott
authors.forename: Noorhan
authors.initials: N
authors.lastname: Chelebi
authors.forename: John
authors.initials: J
authors.lastname: Smith
authors.forename: Tom
authors.initials: T
authors.lastname: Brown
authors.forename: Günter
authors.initials: G
authors.lastname: Schmidt
mesh_terms: D002138:Calibration; D023281:Genomics / Q000379:methods*; D015345:Oligonucleotide Probes / Q000032:analysis* / Q000138:chemical synthesis / Q000737:chemistry; D020135:Peptide Nucleic Acids / Q000032:analysis* / Q000138:chemical synthesis / Q000737:chemistry; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods* / Q000592:standards; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D015345:Oligonucleotide Probes; D020135:Peptide Nucleic Acids