Nonradioactive hybridization probes prepared by the reaction of biotin hydrazide with DNA

“…Several chemical labeling systems have been developed, ranging from post‐synthetic modification of oligonucleotide sequences by phosphoramidite chemistry to photo‐activated reagents such as psoralen 8. Other covalent modifications can be achieved through transamination between cytidine derivatives and primary amines resulting in N 4 ‐substituted cytidines or modification of C 5 of cytidine and uridine by an allylamine and subsequent condensation with activated esters of haptens 9, 10…”

Section: Introductionmentioning

confidence: 99%

Platinum(II)‐Based Coordination Compounds as Nucleic Acid Labeling Reagents: Synthesis, Reactivity, and Applications in Hybridization Assays

Heetebrij

Talman²,

Velzen³

et al. 2003

ChemBioChem

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The synthesis, characterization, and molecular interactions of platinum(II) coordination compounds, which contain a distal nonradioactive reporter molecule, with mono- and polynucleotides are described. A [Pt(II)(en)(NH(2)(CH(2))(6)NH-tBoc)Cl](NO(3)) (en=ethylenediamine) entity has been coupled, after removal of the tBoc group, to a number of hapten and fluorophore molecules through succinimide derivatives. The influence of the various tethered reporter groups within these complexes on the reactivity towards guanosine 5'-monophosphate (5'-GMP), as a model for polynucleotide sequences, was investigated to shed light on the use of these reagents in hybridization assays. Reactivity turned out to be strongly dictated by the chemical nature of the distal reporter molecule present. At pH 7.0 the sequence of reactivity is cationic approximately aromatic (stacking) > neutral > anionic; there is approximately an order of magnitude difference between the fastest reacting complex (k=10.2 x 10(-2) M(-1) s(-1)) and the slowest reacting complex (k=0.93 x 10(-2) M(-1) s(-1)) under these conditions. Platination of an oligodeoxynucleotide (30-mer), dsDNA, or an RNA transcript, shows that a Pt/nucleotide ratio between 1:10 and 1:20 (established by using flameless atomic absorption spectroscopy) results in probes with excellent hybridization characteristics. In terms of applicability and detection limits these platinated nucleic acid probes perform equally well compared to conventionally generated nucleic acid probes, that is, through enzymatic incorporation of covalently labeled nucleotide triphosphates. Applications of these reagents to in situ hybridization assays and gene expression profiling on microarrays illustrate the potential of these monofunctional binding platinum triamine compounds.

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“…A variant of BACH with a shorter spacer arm also exists. It has successfully been used to modify oligonucleotides,24 nerve growth factor,25 low‐density lipoprotein receptor26 and antibodies 27, 28. Shinohara et al 29.…”

mentioning

confidence: 99%

Mass spectrometric analysis of carbohydrates labeled with a biotinylated tag

Kapková¹

2009

Rapid Comm Mass Spectrometry

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A derivatization method for mass spectrometric analysis of oligosaccharides is presented. Small saccharides, complex, high-mannose-type oligosaccharides and oligosaccharides released from hen ovalbumin were converted into their biotin derivatives by incubating them with biotinamidocaproyl hydrazide (BACH). Improved sensitivity of mass spectrometric analysis of labeled glycans in comparison with their natural counterparts was achieved after derivatization. The labeling reagent contains a biotin handle at one end and a hydrazide group at the other. Hence, the key feature of biotinylated sugars is that in addition to their usefulness in functional studies (e.g. analysis of the interaction between lectins and biotin-derivatized oligosaccharides) they might be utilized also for structural analysis of oligosaccharides. Mass spectrometric studies were performed by matrix-assisted laser desorption/ionization time-of-flight and electrospray ionization mass spectrometry.

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Nonradioactive hybridization probes prepared by the reaction of biotin hydrazide with DNA

Cited by 75 publications

References 18 publications

Conjugates of oligonucleotides and modified oligonucleotides: a review of their synthesis and properties

Conjugates of oligonucleotides and modified oligonucleotides: a review of their synthesis and properties

Platinum(II)‐Based Coordination Compounds as Nucleic Acid Labeling Reagents: Synthesis, Reactivity, and Applications in Hybridization Assays

Mass spectrometric analysis of carbohydrates labeled with a biotinylated tag

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