title: Biochemical assays of immobilized oligonucleotides with mass spectrometry.
abstract: This paper reports the use of mass spectrometry to characterize oligonucleotides immobilized to the surfaces of biochips. Biotinylated oligonucleotides were immobilized to self-assembled monolayers that present a streptavidin layer and then treated with a complementary strand to present short duplexes. Treatment of the surface with 5-methoxysalicylic acid and ammonium citrate matrix allows the individual oligonucleotides to be observed by matrix-assisted laser desorption/iozation and time-of-flight mass spectrometry (MALDI-TOF MS). Examples are shown wherein this method is applied to assays of hybridization, of cleavage by a deoxyribozyme, of a dephosphorylation reaction, and of the adducts formed on treatment of DNA with cis-platin. This work provides an early example of the application of mass spectrometry to DNA biochips and may substantially expand the applications of the now common oligonucleotide arrays.
authors.affiliation: Department of Chemistry, Howard Hughes Medical Institute, The University of Chicago, Chicago, Illinois 60637, USA.
authors.forename: Haim
authors.initials: H
authors.lastname: Tsubery
authors.forename: Milan
authors.initials: M
authors.lastname: Mrksich
mesh_terms: D001671:Biochemistry / Q000379:methods*; D001681:Biological Assay; D019921:Biotinylation; D019343:Citric Acid / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D062425:Hydroxybenzoate Ethers; D013058:Mass Spectrometry / Q000379:methods*; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000737:chemistry*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D012459:Salicylates / Q000737:chemistry; D020349:Surface Plasmon Resonance; D013499:Surface Properties
chemical_list: D062425:Hydroxybenzoate Ethers; D009841:Oligonucleotides; D000644:Quaternary Ammonium Compounds; D012459:Salicylates; C033304:5-methoxysalicylic acid; D019343:Citric Acid; C481046:ammonium citrate; D004247:DNA