A simple approach has been developed to prepare conjugates between oligodeoxyribonucleotides (ODNs) and linear copolymers of N-vinylpyrrolidone (NVP)/N-acryloxysuccinimide (NAS). These conjugates were of higher than expected molecular weight, as determined by light scattering. Side reactions between amino groups on heterocyclic bases, especially of cytosine, are proposed to explain the formation of aggregates. ODN-NVP/NAS conjugates used in both capture and detection steps in sandwich hybridization increased assay sensitivity. Sensitivity to 10(7) DNA molecules/mL was reached for a 2.3 kb PCR fragment of hepatitis B virus (HBV) genomic DNA.
SYNOPSISNew azonitrile initiators carrying N-hydroxysuccinimide (ACPS) or biotin derivative (ACP-LC-B) moieties were synthesized from 4,4'-azobis(4-~yanopentanoic acid) (ACPA) with a yield of around 60%. Special care was taken in the assignment of the l3C-NMR resonances of the ACP-LC-B. The kinetic and thermal parameters associated with these initiators were determined by UV spectroscopy and differential scanning calorimetry.
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