Chromophoric reagents for incorporation of biotin or other haptens into macromolecules

• US 5,614,368 A
• Filed: 12/23/1994
• Issued: 03/25/1997
• Est. Priority Date: 02/09/1990
• Status: Expired due to Term

• Current Assignee: Molecular Devices
• Original Assignee: Molecular Devices
• Inventors: Zuk, Robert F., Kung, Viola T., Ghazarossian, Vartan
• Primary Examiner(s): CEPERLEY, MARY
• Application Number:
  US08/363,608
• Time in Patent Office: 823 Days
• Field of Search: 436/501, 436/546, 436/818, 435/7.5
• US Class Current: 435/7.5
• CPC Class Codes: C07D 495/04 Ortho-condensed systems
• CPC Class Codes: G01N 33/532 Production of labelled immunochemicals
• CPC Class Codes: Y10S 436/818 Human chorionic gonadotropin

Abstract

Compounds and methods for detecting the incorporation of a hapten into a protein or macromolecule are disclosed. The compounds comprise haptens bound directly or indirectly to a chromophoric group and a macromolecule-reactive group, whereby incorporation of the hapten into a protein or other macromolecule may be detected.

Claims

1. A method for determining whether a labelled-hapten reagent has been linked to a macromolecule comprising:

(a) contacting a sample containing a macromolecule with a a labelled hapten reagent; and

(b) detecting the label, the labelled hapten reagent having the formula;

##STR10## where x is 4;

y is 1;

n₁ is an integer of from 1-7;

n₂ is 0 or an integer of from 1-7;

R₁ represents a detectable label; and

R₂ is a group which binds directly or indirectly to the macromolecule.

2. A method according to claim 1, wherein Hapten represents ##STR11##

3. A method according to claim 1, wherein the labelled hapten reagent has the formula:

##STR12##

4. A method according to claim 3, wherein the macromolecule-reactive group is an active ester, amino group, sulfhydryl group, carbohydrate group, azido group or a carboxy group.

5. A method according to claim 3, wherein the macromolecule-reactive group is an N-hydroxysuccinimide ester or an N-hydroxysulfosuccinimide ester.

6. A method according to claim 1, wherein the detectable label is carboxytetramethyl rhodamyl, sulforhodamyl 101, or dinitrophenylaminohexanoyl.

7. A method according to claim 1, wherein R₁ is carboxytetramethyl-rhodamyl, sulforhodamyl 101, or dinitrophenylaminohexanoyl;

and R₂ is --CO₂ H, --NH₂, --SH, ##STR13##