title: A feasible approach to evaluate the relative reactivity of NHS-ester activated group with primary amine-derivatized DNA analogue and non-derivatized impurity.
abstract: Synthetic DNA analogues with improved stability are widely used in life science. The 3'and/or 5' equivalent terminuses are often derivatized by attaching an active group for further modification, but a certain amount of non-derivatized impurity often remains. It is important to know to what extent the impurity would influence further modification. The reaction of an NHS ester with primary amine is one of the most widely used options to modify DNA analogues. In this short communication, a 3'-(NH2-biotin)-derivatized morpholino DNA analogue (MORF) was utilized as the model derivatized DNA analogue. Inclusion of a biotin concomitant with the primary amine at the 3'-terminus allows for the use of streptavidin to discriminate between the products from the derivatized MORF and non-derivatized MORF impurity. To detect the MORF reaction with NHS ester, S-acetyl NHS-MAG3 was conjugated to the DNA analogue for labeling with (99m)Tc, a widely used nuclide in the clinic. It was found that the non-derivatized MORF also reacted with the S-acetyl NHS-MAG3. Radiolabeling of the product yielded an equally high labeling efficiency. Nevertheless, streptavidin binding indicated that under the conditions of this investigation, the non-derivatized MORF was five times less reactive than the amine-derivatized MORF.
authors.affiliation: a Department of Radiology , University of Massachusetts Medical School , Worcester , Massachusetts.
authors.forename: Shuping
authors.initials: S
authors.lastname: Dou
authors.forename: John
authors.initials: J
authors.lastname: Virostko
authors.forename: Dale L
authors.initials: DL
authors.lastname: Greiner
authors.forename: Alvin C
authors.initials: AC
authors.lastname: Powers
authors.forename: Guozheng
authors.initials: G
authors.lastname: Liu
mesh_terms: D000588:Amines / Q000737:chemistry*; D001710:Biotin / Q000737:chemistry; D002614:Chelating Agents / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000737:chemistry*; D004952:Esters / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D009025:Morpholines / Q000737:chemistry; D009842:Oligopeptides / Q000737:chemistry; D019809:Streptavidin / Q000737:chemistry; D013388:Succinimides / Q000737:chemistry; D013667:Technetium / Q000737:chemistry
chemical_list: D000588:Amines; D002614:Chelating Agents; D004952:Esters; D009025:Morpholines; D009842:Oligopeptides; C118517:S-acetylmercaptoacetyltriglycine N-hydroxysuccinimide; D013388:Succinimides; D001710:Biotin; D013667:Technetium; D004247:DNA; D019809:Streptavidin
keywords: Chelator; DNA analogues; conjugation; radiolabeling