Phenethylamine antigenic conjugates, their preparation, antibodies and use

Claim 2

A synthetic antigen of formula: ##STR10## where R' and R" of the phenethylamine hapten are individually selected from the class consisting of --H, --OH, and --OCH 3 , where when either R' or R" is --OCH 3 the other is --OH; R'" of the phenethylamine hapten is selected from the class consisting of --H and --OH; R"" of the phenethylamine hapten is selected from the class consisting of --H, --OCH 3 , and --C 2 H 5 ; Q of the phenethylamine hapten is selected from the class consisting of --H and --CH 3 ; Y is a linking group which is the residue of the reaction of a coupling agent with a reactive group derivatized into the phenol ring and of the coupling agent with the carrier; and the carrier is a macromolecule conferring antigenicity.

Claim 3

The synthetic antigen of claim 1 wherein Y is diazo and R' and R" are both --H.

Claim 4

The synthetic antigen of claim 1 in which Y is a dicarboxylic acid residue.

Claim 5

The synthetic antigen of claim 1 wherein Y includes a phenazo group bonded to the phenethylamine ring and the phenethylamine includes an aromatic hydroxyl.

Claim 6

Protein-p-diazoamphetamine.

Claim 7

Protein-p-diazoamphetamine.

Claim 8

Protein-p-succinylamphetamine.

Claim 9

Protein-p-succinylamphetamine.

Claim 10

Protein-p-benzoyldiazo-m-epinephrine.

Claim 11

Protein-p-benzoyldiazo-m-epinephrine.

Claim 12

Protein-p-benzoyldiazo-p-norphenylephrine

Claim 13

Protein-p-benzoyldiazo-p-norphenylephrine

Claim 14

Antibody raised by the antigen of claim 1 and specific to the phenethylamine hapten.

Claim 15

Antibody raised by the antigen of claim 5 and specific to the amphetamine.

Claim 16

Antibody raised by the antigen of claim 6 and specific to amphetamine.

Claim 17

Antibody raised by the antigen of claim 7 and specific to the epinephrine.

Claim 18

Antibody raised by the antige of claim 8 and specific to norphenylephrine.

Claim 19

The antibody of claim 9 insolubilized by securing it to a solid matrix.

Claim 20

The antibody of claim 10 insolubilized by securing it to a solid matrix.

Claim 21

The antibody of claim 11 insolubilized by securing it to a solid matrix.

Claim 22

The antibody of claim 12 insolubilized by securing it to a solid matrix.

Claim 23

The antibody of claim 13 insolubilized by securing it to a solid matrix.

Claim 24

An immunochemical method for assaying for the presence of an antigen target in a sample, said antigen having the formula: ##STR11## where R' and R" are individually selected from the class consisting of --H, --OH, and --OCH 3 , where when either R' and R" is --OCH 3 the other is --OH; R'" is selected from the class consisting of --H and --OH; R"" is selected from the class consisting of --H, --OCH 3 , and --C 2 H 5 ; Q is selected from the class consisting of --H and --CH 3 ; wherein said method employs an antibody obtained by the immunologic response of a vertebrate animal to administration of an antigen according to claim 1 and wherein said antibody is specific to the target, said method also employing a standard, the antibody binding with the target to form an antibody-target complex and competitively binding with the standard to form an antibody-standard complex, the antibodystandard complex having an artificially introduced radiation label enabling the complex to be assayed quantitatively by measurement of radiation emanating from it, the affinities of the antibody for the standard and for the target being known quantitatively, said method comprising allowing a known quantity of the sample and a known quantity of the standard to compete for binding with a known quantity of the antibody and determining the radiation emanating from the antibody-standard complex, thereby enabling the quantity of antibody-bound standard to be calculated and the quantity of target in the sample to be deduced.

Claim 25

The method of claim 19 wherein the label is radioactive and the antibody-standard complex is separated from any non-complexed labeled material after allowing competition binding and before determination of the emanated radiation.

Claim 26

The method of claim 19 wherein the label is fluorescent and the standard is provided with a chemical moiety giving it a fluorescence spectrum overlapping the natural fluorescence spectrum of the antibody, whereby the complex can be assayed by measurement of the perturbation of the antibody fluorescence due to its binding with the standard.