Visualization polymers and their application to diagnostic medicine

A detection kit for analysis of target molecules in a biological material comprising metered quantities of a buffered, aqueous solution of a detection agent component and a buffered, aqueous solution of a visualization polymer complex component and a standardized quantity of an aqueous mixture of said visualization polymer, said detection agent component and said visualization polymer complex component being capable of associating when combined,

• said visualization polymer complex component comprising a visualization polymer covalently bonded to a ligand, and a ligand binding compound associated with said ligand;
• said visualization polymer comprising essentially identical multiple visualization units each having at least one visualization site;
• said visualization units being directly bonded together through chemical groups or backbone moieties of said visualization units, or linked by a coupling agent covalently bonded to chemical groups or backbone moieties of said visualization units;
• said visualization unit being an enzyme, wherein the enzymatic site is the visualization site, a tagged natural or synthetic polypeptide, a tagged polyol, a tagged polyolefin or a tagged polycarbohydrate wherein the tag of the tagged polypeptide, tagged polyol, tagged polyolefin or tagged polycarbohydrate is the visualization site thereof,
• said chemical group being an amine group, an oxidized 1,2-diol group, a carboxy group, a mercaptan group, a hydroxy group or a carbon-hydrogen bond;
• said backbone moiety being an amide bond, a carbon-carbon bond, a carbon-oxygen bond or a carbon-hydrogen bond;
• said chemical group or backbone moiety being located at a position which is at least one atom away from the visualization site of said visualization unit;
• said coupling agent being derived from a bi- or multi-(homo-or hetero-) functional organic cross-linking reagent of the formula ##STR5## wherein A, B and E are independently selected from the group consisting of hydrogen, a carboxylic acid group, an acid halide group, an activated ester, a mixed anhydride, an acyl imidazole, an N-(carbonyloxy)imide group, an iminoester, a primary amine, an aldehyde group, an alphahalomethylcarbonyl group, a hydrazine group, an acylhydrazide group, an azide group and an N-maleimide group;
• at least two of A, B and E are other than hydrogen;
• and R.sup.1 is an aliphatic group of at least two carbons or an aromatic group of at least six carbons;
• and
• the tag of said tagged polypeptide, tagged polyol, tagged polyolefin or tagged carbohydrate is a fluorescent group, a dye, a radioactive group, a photon emitter or an electron dense group.

A method for visualizing the presence of an inorganic or organic target molecule in a biological material, which comprises:

• combining said target with a detecting agent with carries a visualization polymer comprising essentially identical multiple visualization units directly bonded together or linked by a coupling agent through chemical groups or backbone moieties of said visualization units;
• said visualization unit having at least one visualization site and said visualization unit being an enzyme wherein the enzymatic site is the visualization site, a tagged natural or synthetic polypeptide, a tagged polyol, a tagged polyolefin or a tagged polycarbohydrate wherein the tag of the tagged polypeptide, the tagged polyol, the tagged polyolefin or the tagged polycarbohydrate is the visualization site thereof,
• said chemical group being an amine group, an oxidized 1,2-diol group, a carboxy group, a mercaptan group, a hydroxy group or a carbon-hydrogen bond;
• said chemical group or backbone moiety being located within said visualization unit at a position which is at least one atom away from the visualization site of said visualization unit;
• said coupling agent being derived from a reactive bi- or multi- homo- or hetero-functional organic cross-linking reagent of the formula ##STR6## wherein A, B and E are independently selected from the group consisting of hydrogen, a carboxylic acid group, an acid halide group, an activated ester, a mixed anhydride, an acyl imidazole, an N-(carbonyloxy)imide group, an iminoester, a primary amine, an aldehyde group, an alphahalomethylcarbonyl group, a hydrazine group, an acylhydrazide group, an azide group and an N-maleimide group;
• at least two of A, B and E are other than hydrogen;
• and R.sup.1 is an aliphatic group of at least two carbons or an aromatic group of at least six carbons;
• and
• the tag of the tagged polypeptide, tagged polyol, tagged polyolefin or tagged carbohydrate is a fluorescent chemical group, a dye, a radioactive group, a photon emitter or an electron dense moiety.