5-Fluoro-uracil immunoassay
Novel conjugates of 5-fluoro-uracil and novel 5-fluoro-uracil immunogens and monoclonal antibodies generated by these immunogens which are useful in immunoassays for the quantification and monitoring of 5-fluoro-uracil in biological fluids.
1 . An immunoassay for detecting 5-fluoro-uracil in a sample providing a mixture of a sample, containing an antibody selectively reactive with 5-fluoro-uracil and not substantially cross-reactive with tegafur and a conjugate of a carrier with a compound of the formula:
wherein Y is an organic spacing group;
X is a terminal functional group capable of binding to a polyamine polymer; and
p is an integer from 0 to 1;
causing the 5-fluoro-uracil in the sample and said conjugate to bind with said antibody and thereafter measuring the amount of said conjugate in said mixture which is bound or unbound to said antibody whereby the presence of 5-fluoro-uracil in the sample can be determined.
2 . The process of claim 1 , wherein the sample is a human sample.
3 . The immunoassay of claim 2 , wherein said antibody is generated from an immunogen comprising an immunogenic polymer linked to a compound of the formula:
wherein p, X and Y are as above.
4 . The immunoassay of claim 2 , wherein the antibody is attached to a solid support.
5 . The immunoassay of claim 4 , wherein the solid support is microtitor plates.
6 . The immunoassay of claim 4 , wherein the solid support is nanoparticles.
7 . An antibody which binds selectively to 5-fluoro-uracil and does not substantially bind to tegafur.
8 . The antibody of claim 8 , wherein said antibody is a monoclonal antibody.
9 . The antibody of claim 8 , wherein said antibody is derived from mice, rabbits or rats.
10 . The antibody of claim 9 , wherein said antibody does not substantially cross-react with uracil and cytosine.
11 . The antibody of claim 7 , wherein said antibody is derived from an immunogen of a polyamine polymer with a compound of the formula:
wherein Y is an organic spacing group;
X is a terminal functional group capable of binding to a polyamine polymer; and
p is an integer from 0 to 1.
12 . The antibody of claim 11 , wherein said antibody is a monoclonal antibody.
13 . The antibody of claim 11 , wherein said antibody is derived from mice, rabbits or rats.
14 . The antibody of claim 13 , wherein said antibody does not substantially cross-react with uracil and cytosine.
15 . A compound of the formula:
wherein Y is an organic spacing group;
X is a terminal functional group capable of binding to a polyamine polymer; and
p is an integer from 0 to 1.
16 . The compound of claim 15 , wherein p is 0.
17 . The compound of claim 16 , wherein X is
wherein R 3 is hydrogen or taken together with its attached oxygen atom forms a reactive ester and R 4 is oxygen or sulfur.
18 . The compound of claim 17 , wherein
and R 3 is hydrogen.
19 . The compound of claim 17 , wherein
and R 3 forms a reactive ester.
20 . The compound of claim 19 , wherein the ester formed is a lower alkyl ester, imidoester or amidoester.
21 . The compound of claim 15 , wherein p is 1.
22 . The compound of claim 21 , wherein Y is alkylene containing from 1 to 10 carbon atoms,
wherein n and o are integers from 0 to 6, and m is an integer from 1 to 6.
23 . A compound of the formula:
wherein p, X and Y are as above.
24 . The compound of claim 23 , wherein p is 0.
25 . The compound of claim 24 , wherein X is
wherein R 3 is hydrogen or taken together with its attached oxygen atom forms a reactive ester and R 4 is oxygen or sulfur.
26 . The compound of claim 25 , wherein
and R 3 is hydrogen.
27 . The compound of claim 26 , wherein
and R 3 forms a reactive ester.
28 . The compound of claim 27 , wherein the ester formed is a lower alkyl ester, imidoester or amidoester.
29 . The compound of claim 23 , wherein p is 1.
30 . The compound of claim 29 , wherein X is
wherein R 3 is hydrogen or taken together with its attached oxygen atom forms a reactive ester and R 4 is oxygen or sulfur.
31 . The compound of claim 30 , wherein Y is alkylene containing from 1 to 10 carbon atoms,
wherein n and o are integers from 0 to 6, and m is an integer from 1 to 6.
32 . A conjugate of a carrier with a compound of the formula:
wherein Y is an organic spacing group;
X is a terminal functional group capable of binding to a polyamine polymer; and
p is an integer from 0 to 1.
33 . The conjugate of claim 32 , wherein p is 0.
34 . The conjugate of claim 33 , wherein X is
wherein R 3 is hydrogen or taken together with its attached oxygen atom forms a reactive ester and R 4 is oxygen or sulfur.
35 . The conjugate of claim 32 , wherein p is 1 and Y is alkylene containing from 1 to 10 carbon atoms,
wherein n and o are integers from 0 to 6, and m is an integer from 1 to 6.
36 . The conjugate of claim 35 , wherein the carrier contains one or more amino groups linked by
wherein R 4 is oxygen or sulfur.
37 . A compound of the formula:
wherein p, X and Y are as above.
38 . The compound of claim 37 , wherein p is 0.
39 . The compound of claim 38 , wherein X is
wherein R 3 is hydrogen or taken together with its attached oxygen atom forms a reactive ester and R 4 is oxygen or sulfur.
40 . The compound of claim 37 , wherein p is 1.
41 . The compound of claim 40 , wherein Y is alkylene containing from 1 to 10 carbon atoms,
wherein n and o are integers from 0 to 6, and m is an integer from 1 to 6.
42 . The compound of claim 41 , wherein X is
wherein R 3 is hydrogen or taken together with its attached oxygen atom forms a reactive ester and R 4 is oxygen or sulfur.
43 . An immunogen containing an immunogenic polyamine polymer linked to a compound of the formula:
wherein Y is an organic spacing group;
X is a terminal functional group capable of binding to a polyamine polymer; and
p is an integer from 0 to 1.
44 . The immunogen of claim 43 , wherein p is 0.
45 . The immunogen of claim 43 , wherein p is 1.
46 . The immunogen of claim 45 , wherein Y is alkylene containing from 1 to lo carbon atoms,
wherein n and o are integers from 0 to 6, and m is an integer from 1 to 6.
47 . The immunogen of claim 46 , wherein the immunogenic polymer contains one or more amino groups linked by
wherein R 4 is oxygen or sulfur.
48 . A kit for determining the presence of 5-fluoro-uracil in a patient sample comprising reagents in separate containers, one of the reagents being a conjugate of a carrier with a compound of the formula:
wherein Y is an organic spacing group;
X is a terminal functional group capable of binding to a polyamine polymer; and
p is an integer from 0 to 1
and the second container containing an antibody substantially selectively reactive with 5-fluoro-uracil and not substantially cross-reactive to tegafur or uracil.
49 . The kit of claim 48 , wherein said conjugate is present in a predetermined amount in said first container.
50 . The kit of claim 49 , wherein said kit is used to determine the amount of 5-fluoro-uracil in said sample.
51 . The kit of claim 48 , wherein, said antibody is generated from an immunogen of an immunogenic polyamine polypeptide linked to a compound of the formula:
wherein p, X and Y are as above.