US 6,190,873 B1 · Cyclosporin assay and kit

Abstract

A method of measuring the amount of cyclosporin in a sample suspected of containing cyclosporin is disclosed. A method of inactivating interfering cross-reactive material in an assay for measuring the amount of cyclosporin in a sample suspected of containing cyclosporin is also disclosed. Compositions wherein cyclosporin is conjugated to an immunogenic carrier or a label, optionally through a linking group, at an alanine nitrogen atom of the cyclic backbone of cyclosporin are also disclosed. Compositions wherein atiocyclosporin is conjugated, optionally through a linking group, to an immunogenic carrier or a label are also disclosed. Where cyclosporin is conjugated to an immunogenic carrier, the conjugates may be used as immunogens for the preparation of antibodies which are capable of recognizing cyclosporin. Where atiocyclosporin is conjugated to an immunogenic carrier, the conjugates may be used as immunogens for the preparation of antibodies which are capable of recognizing interfering cross-reactive material but substantially incapable of recognizing cyclosporin or cyclosporin-label conjugates. Where cyclosporin is conjugated to a label, the conjugates may be used as part of a signal producing system in cyclosporin assays. Both the antibodies and label conjugates are useful in the disclosed assay methods.

Claims57

Dependent claims

An assay for the presence of cyclosporin in a sample suspected of containing cyclosporin, wherein the assay comprises:

(A) combining in an aqueous medium:
1) said sample suspected of containing cyclosporin,
2) a cyclosporin-label conjugate comprising cyclosporin conjugated at an alanine nitrogen atom of amino acid residue 7 or 8 to a label, and
3) antibodies capable of binding to said cyclosporin-label conjugate, wherein said antibodies have been raised against a cyclosporin-immunogenic carrier conjugate comprising cyclosporin conjugated to an immunogenic carrier,
(B) determining the presence of a detectable complex;
and
(C) correlating said detectable complex with the presence of cyclosporin in said sample;
wherein said cyclosporin is conjugated to said label and said immunogenic carrier through a first and a second linking group, respectively;
wherein said linking groups are independently selected from the group consisting of
(I) a hydroxyalkane chain of the formula —(CH2)nO—, where n is an integer of about 1 to 6;
(II) a carboxyl benzyl group of the formula:
wherein an alanine nitrogen atom of cyclosporin is conjugated to the benzylic carbon atom in the carboxyl benzyl group;
and
(III) a moiety of the formula:

11 more elements

Dependencies are inferred from the claim text.

Cyclosporin assay and kit

Life of this patent

Cyclosporin assay and kit

Life of this patent

Abstract

Claims57