Claims
- 1. An inactivated avidin or streptavidin of formula II ##STR7## wherein R=�(CH.sub.2).sub.n --O.sub.m !.sub.q --�(CH.sub.2).sub.r --O.sub.s !.sub.t --(CH.sub.2).sub.p, wherein n, r=2, 3 or 4; m,s=1; q+t=1-4; and p=1-4, and the sum of all CH.sub.2 groups in R does not exceed 10; and
- R.sub.1 is at least one ring substituent selected from the group consisting of hydrogen, C.sub.1 -C.sub.5 alkyl, NH.sub.2, COOH, NO.sub.2, F, Cl and Br, produced by a process comprising:
- (a) providing a photo-activatable biotin derivative of formula I ##STR8## wherein R and R.sub.1 are as defined above; and (b) contacting the biotin derivative with avidin or streptavidin ##STR9## wherein " ##STR10## " represents the active center of the avidin or streptavidin, and triggering a photoreaction, to specifically bind the biotin derivative to the active center of the avidin or streptavidin via the ##STR11## group, and to covalently bind the biotin derivative to the avidin or streptavidin outside the active center via the photo-activatable N3 group.
- 2. A complex comprising avidin or streptavidin having bound thereto a photo-activatable biotin derivative, wherein the complex is of formula II ##STR12## wherein R=�(CH.sub.2).sub.n --O.sub.m !.sub.q --�(CH.sub.2).sub.r --O.sub.s !.sub.t --(CH.sub.2).sub.p, wherein n, r=2, 3 or 4; m,s=1; q+t=1-4; and p=1-4, and the sum of all CH.sub.2 groups in R does not exceed 10,
- R.sub.1 is at least one ring substituent selected from the group consisting of hydrogen, C.sub.1 -C.sub.5 alkyl, NH.sub.2, COOH, NO.sub.2, F, Cl and Br, and " ##STR13## " represents the active center of the avidin or streptavidin which is specifically bound by the biotin derivative via the ##STR14## group, and wherein the biotin derivative is covalently bound to the avidin or streptavidin outside the active center via a photo-activatable group.
- 3. A method for the immunological determination of an analyte in a sample comprising the following steps:
- a) contacting the sample which may contain the analyte with inactivated streptavidin or avidin of claim 1;
- b) contacting the sample with at least one specific binding partner of the analyte to form a complex, wherein step a) and step b) can occur in any order or simultaneously; and
- c) measuring the formed complex of analyte and specific binding partners as a measure of the presence of the analyte.
- 4. A method of reducing interference in an immunoassay which requires the formation of a complex between an analyte and a specific binding partner of the analyte, comprising conducting the immunoassay in the presence of the inactivated streptavidin or avidin of claim 1.
- 5. A method for the immunological determination of an analyte in a sample comprising the following steps:
- a) contacting the sample which may contain the analyte with inactivated streptavidin or avidin of claim 2;
- b) contacting the sample with at least one specific binding partner of the analyte to form a complex, wherein step a) and step b) can occur in any order or simultaneously; and
- c) measuring the formed complex of analyte and specific binding partners as a measure of the presence of the analyte.
- 6. A method of reducing interference in an immunoassay which requires the formation of a complex between ah analyte and a specific binding partner of the analyte, comprising conducting the immunoassay in the presence of the inactivated streptavidin or avidin of claim 2.
- 7. The method of claim 3, wherein the immunoassay further comprises the use of the specific binding pair avidin/biotin or streptavidin/biotin.
- 8. The method of claim 5, wherein the immunoassay further comprises the use of the specific binding pair avidin/biotin or streptavidin/biotin.
- 9. The method of claim 4, wherein streptavidin or avidin is also used as a binding partner in the immunoassay.
- 10. The method of claim 6, wherein streptavidin or avidin is also used as a binding partner in the immunoassay.
- 11. Test kit for the determination of an analyte comprising, in separate containers:
- a) inactivated streptavidin or avidin of claim 2, and
- b) at least one specific binding partner of the analyte.
- 12. A photo-activatable biotin derivative of formula I ##STR15## wherein R=--�(CH.sub.2).sub.n --O.sub.m !.sub.q --�(CH.sub.2).sub.r --O.sub.s !.sub.t --(CH.sub.2).sub.p--
- wherein n and r are each independently=2, 3 or 4; m and s are each independently=1; q+t=1-4; and p=14;
- wherein the sum of all CH.sub.2 groups in R does not exceed 10;
- and wherein R.sub.1 is at least one ring substituent selected from the group consisting of hydrogen, C.sub.1 -C.sub.5 -alkyl, NH.sub.2, COOH, F, Cl, and Br.
- 13. The biotin derivative of claim 12, wherein n=3, r=4, m,s=1, q,t=1, p=3.
- 14. The biotin derivative of claim 12, wherein n,r=2, m,s=1, q,t=1, p=2.
- 15. Test kit for the determination of an analyte comprising, in separate containers:
- a) inactivated streptavidin or avidin of claim 1, and
- b) at least one specific binding partner of the analyte.
- 16. A method for the preparation of a photo-activatable biotin derivative of claim 12, comprising:
- a) attaching a diamino compound of the formula III
- NH.sub.2 --R--NH.sub.2 (III)
- wherein R=�(CH.sub.2).sub.n --O.sub.m !.sub.q --�(CH.sub.2).sub.r --O.sub.s !.sub.t --(CH.sub.2).sub.p
- wherein n,r=2,3; m,s=1; q+t=1-4; p=1-4;
- wherein the sum of all CH.sub.2 groups in R does not exceed 10;
- 1) at one end to the carboxyl group of biotin and
- 2) at the other end to a photo-activatable group having the formula ##STR16## wherein R.sub.1 is at least one ring substituent selected from the group consisting of hydrogen, C.sub.1 -C.sub.5 -alkyl, NH.sub.2, COOH, F, Cl, and Br.
- 17. A method for inactivating streptavidin or avidin with a photo-activatable biotin derivative comprising:
- a) contacting the avidin or streptavidin with the biotin derivative, and
- b) coupling the avidin or streptavidin and the biotin by exposure to radiation,
- wherein the biotin derivative is a compound of claim 12.
Priority Claims (1)
Number |
Date |
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44 07 423 |
Mar 1994 |
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Parent Case Info
This application is a continuation of application Ser. No. 08/535,072, filed Nov. 3, 1995 now abandoned which is a 371 of PCT/EP95/00690, filed Feb. 25, 1995.
US Referenced Citations (1)
Number |
Name |
Date |
Kind |
5395938 |
Ramakrishnan |
Mar 1995 |
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Foreign Referenced Citations (3)
Number |
Date |
Country |
155854 |
Sep 1985 |
EPX |
331068 |
Sep 1989 |
EPX |
4302241 |
Jul 1994 |
DEX |
Non-Patent Literature Citations (1)
Entry |
U. Henriksen et al., J. Photochemistry and Photobiology, vol. 57, pp. 331-342, 1991. |
Continuations (1)
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Number |
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Parent |
535072 |
Nov 1995 |
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