Tyrosyl-class antigenic conjugates, their preparation and antibodies raised thereto

A process for preparing an antigen of the formula:

• ##SPC6##
• wherein the carrier is selected from the class consisting of proteins and synthetic proteins, and confers antigenicity, R'" is coupled to the carrier through a reactive group, the residue being substantially non-reactive R'" being bonded to the aryl with up to three bonds, being substituted or unsubstituted, and having up to four rings and up to 20 aliphatic carbon atoms;
• and the hapten has the formula:
• ##STR3## where R' is selected from the group consisting of -OH and -NHP.sub.p.sub.' and R" is selected from the class consisting of --H and --COP.sub.p.sub.' where P.sub.p is a polypeptide chain having from 1 to 250 peptide groups, with or without pendant groups;
• X is selected from the class consisting of H and I;
• X' is selected from the class consisting of H and I when X is I;
• and n is 0 or 1 and X' is H when n is 0;
• comprising:
• a. conjugation of a primary aromatic amine with a carrier;
• b. separation of the reactants from step (a);
• c. lyophilization of the residue to remove the reaction medium;
• d. diazotization of the residue;
• and,
• e. coupling of the hapten to the resulting product.

A method for forming a synthetic antigen from a carrier selected from the class consisting of proteins and synthetic proteins, said carrier having an available reactive group for coupling thereto, a hapten having a reactive group selected from the class consisting of amino and carboxyl, and a carbon atom susceptible to diazonium coupling, and a primary aromatic amine compound having a reactive residue capable of coupling to the reactive group on the carrier, wherein the carrier reactive group and the primary amine compound reactive residue consist of a carboxyl group and an amino group, whereby the carrier can be coupled to the primary amine compound by an amide group, the method comprising:

• a. conjugation of the primary aromatic amine compound with the carrier to form an amide bond;
• b. treatment of the conjugated primary aromatic amine compound to provide for bonding of the hapten by formation of diazonium ion;
• and
• c. coupling of the hapten to the conjugated diazonium ion.