| 摘要 |
<p>This invention relates to thiol-reactive polynucleotide compounds and processes for use of these compounds to covalently conjugate an enzyme label to an oligonucleotide. Oligonucleotides are made thiol-reactive through a chemical modification which comprises the addition of a primary amine. The oligonucleotide so modified is then reacted with a hetero-bifunctional reagent in an acylation reaction, the hetero-bifunctional reagent having an amino-reactive and a thiol-reactive portion, such as an N-hydroxy succinimidyl (NHS) ester and an α-bromoacetamide, respectively. The enzyme is derivatized with a blocked, thiol-containing amino reactive reagent, such as dithio-bis-propionic acid N-hydroxysuccinimide ester. Conjugation is achieved under mild physiologic conditions through a specific reaction of the reactive moieties.</p> |
