C - Chemistry – Metallurgy – 07 – H
Patent
C - Chemistry, Metallurgy
07
H
260/210, 260/229
C07H 19/04 (2006.01) C07H 21/00 (2006.01) C07H 23/00 (2006.01)
Patent
CA 1194862
ABSTRACT A method for preparing a substrate capable of under- going catalytic-induced hydrolysis of the phosphate ester at the 3'-position to yield a species capable of being monitored spectrophotometrically or fluorometrically and thus useful in carrying out various non-isotopic immunoassays, comprises (a) blocking a mononucleosids of the formula: Image wherein B is a nucleotide base, and wherein the CH2OH group at the 4'-position is either cis or trans to said base, with a silyl blocking member at both the 2'- and 5'-hydroxyls of said mononucleoside to form a 2'-, 5'-diblocked mononucleoside; and (b) forming a 2'-, 5'-diblocked mononucleotide 3'- phosphodiester by bonding said 2'-, 5'-diblocked mononucleoside with a moiety selected from the group consisting of a chromophore or fluorophore; said silyl blocking member at the 2'-hydroxyl being capable of at least essentially blocking medium-induced hydro- lysis of the phosphodiester at the 3'-position, and said silyl blocking member at least at the 2'-hydroxyl being capable of being removed to provide a substrate characterized by the abil- ity to undergo catalytic-induced hydrolysis of the phosphate ester at the 3'-position to yield a species capable of being monitored spectrophotometrically or fluorometrically. The silyl blocking member at least at the 2'-hydroxyl is removed so as to provide a substrate characterized by the ability to undergo cat- alytic induced hydrolysis of said phosphodiester to yield a species capable of being monitored spectrophotometrically or fluorometrically.
397670
Osler Hoskin & Harcourt Llp
Roche Diagnostics Corporation
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