IMMUNOLOGICAL REAGENT AND RADIOIMMUNO ASSAY

• US 3,853,987 A
• Filed: 09/01/1971
• Issued: 12/10/1974
• Est. Priority Date: 09/01/1971
• Status: Expired due to Term

• Current Assignee: William J. Dreyer
• Original Assignee: William J. Dreyer
• Inventors: Dreyer, William J.
• Primary Examiner(s): Padgett, Benjamin R.
• Application Number:
  US05/177,017
• Time in Patent Office: 1,196 Days
• Field of Search: 424/1,8,12 23/23B 252/31.1R 250/303
• US Class Current: 424/1.37
• CPC Class Codes: G01N 33/54313 the carrier being characterised by its particulate form
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine

Abstract

A broad new class of reagents permits extremely sensitive and specific assay for, or chemical separation of, a broad range of biological and nonbiological substances. Each reagent consists of a suspension of microscopic carrier material particles, each particle bearing (1) tracer material-fluorescent, radioactive or otherwise - and (2) a coating of biological antibody for the substance whose assay is desired. The latter substance if introduced into the suspension links the particles together in pairs or clumps, which may be sensitively and accurately detected by monitoring the tracer. The carrier is preferably partially hydrolyzed polyacrylamide resin, or in appropriate applications acrylic acid and other derivatives thereof, and other polymers including agar, and the coupling effected by covalent bonding. Other embodiments, including various mechanical forms of carrier, for greater ease of handling and separation, are also described.

Claims

1. THE PROCESS OF FORMING A REAGENT, THAT INCLUDES:

A. PROVIDING MULTIPLE WATER SUSPENDIBLE POLYMERIC PARTICLES, AND LABELLING EACH PARTICLE GENERALLY THROUGHOUT THE VOLUME WITH A MULTIPLICITY OF TRACER MOLECULES, AND B. COVALENTLY BONDING TO EACH PARTICLE A MULTIPLICITY OF MOLECULES OF A SUBSTANCE SELECTED FROM THE GROUP CONSISTING OF ANTIBODIES, ANTIGENS AND PORTIONS AND EQUIVALENTS THEREOF, BY CONTACTING THE PARTICLES WITH SAID MOLECULES IN AQUEOUS SOLUTION IN WHICH THE PARTICLES ARE DISPERSED.

2. The reagent formed by the process of claim 1.

3. The process of claim 1 wherein the particles are porous to the tracer molecules.

4. The process that employs the reagent of claim 1 in assaying an aqueous sample for a particular reactant selected from the group consisting of antibodies, antigens, portions and equivalents thereof, the substance being the immunological homologue to the reactant, that includes the steps c. mixing the reagent with the aqueous sample to produce linking of multiple particles with units of reactant, thereby to form agglomerate, and d. detecting tracer concentrations in the agglomerate to thereby determine the amount of reactant in the sample.

5. The process that employs the rEagent of claim 1 in assaying an aqueous sample for a particular reactant selected from the group consisting of antibodies, antigens, portions and equivalents thereof, the substance being the immunological homologue to the reactant, that includes the steps c. mixing the reagent with the aqueous sample to produce linking of multiple particles with units of reactant, thereby to form agglomerate, and d. detecting tracer concentrations in a non-agglomerated portion of the mix.

6. The process of claim 1 wherein the tracer molecules are selected from the group consisting of radioactive and fluorescent molecules.

7. The process of claim 1 wherein said particles are selected from the group consisting of polymers of acrylic acid, acrylamide, other acrylic acid derivatives, polymers of styrene and derivatives thereof, agar, agarose, and cellulose acetate.

8. The agglomerate formed by the process that employs the reagent of claim 1 in assaying an aqueous sample for a particular reactant selected from the group consisting of antibodies and antigens, the substance being the immunological homologue to the reactant, and that includes the steps of mixing the reagent with the aqueous sample, and continuing said mixing to produce linking of multiple particles with each unit of reactant, thereby to form the agglomerate.