Measuring reagent kit for determining an antigen, antibody or antigen-antibody complex

• US 4,690,908 A
• Filed: 08/20/1984
• Issued: 09/01/1987
• Est. Priority Date: 06/05/1979
• Status: Expired due to Fees

• Current Assignee: Mochida Seiyaku Kabushiki Kaisha
• Original Assignee: Mochida Seiyaku Kabushiki Kaisha
• Inventors: Shinkai, Hiroyuki, Sugawara, Toshiyuki, Kudo, Takashi, Mochida, Ei
• Primary Examiner(s): Kepplinger, Esther M.
• Assistant Examiner(s): Saunders, David A.
• Application Number:
  US06/642,734
• Time in Patent Office: 1,107 Days
• Field of Search: 436/500, 436/501, 436/507, 436/508, 436/518, 436/517, 436/523, 436/528, 436/529, 436/530, 436/531-534, 436/536, 436/808, 436/826, 436/519, 436/520, 436/524
• US Class Current: 436/518
• CPC Class Codes: G01N 33/54346 Nanoparticles
• CPC Class Codes: Y10S 436/813 Cancer
• CPC Class Codes: Y10S 436/817 Steroids or hormones
• CPC Class Codes: Y10S 436/818 Human chorionic gonadotropin
• CPC Class Codes: Y10S 436/826 Additives, e.g. buffers, diluents, preservatives

Abstract

A method for the quantification of an antigen, antibody, or antigen-antibody complex in a sample solution involving measuring the results of an immunochemical agglutination reaction or an agglutination inhibition reaction by spectrophotometry, wherein after the initiation of the agglutination between the antigen, antibody, or antigen-antibody complex to be determined and sensitized carrier particles to which a substance specifically bindable to the said antigen, antibody, or antigen-antibody complex is bound, a fixing compound is added to fix aggregates formed by the agglutination. A measuring reagent kit or pack includes all the components for use in carrying out the above method.

Claims

1. A measuring reagent kit for an immuno-chemical agglutination reaction measurement of a quantity or concentration of a substance to be measured selected from the group consisting of an antigen, an antibody, and an antigen-antibody complex in a sample, said kit comprising:

a mercantile container, said mercantile container containing at least first, second and third individual containers;

(a) at least one said first individual container containing a reagent consisting essentially of a suspension of sensitized carrier particles for causing agglutination by reaction with the substance to be measured, said particles being formed by binding at least one substance selected from the group consisting of antigens, antibodies, and antigen-antibody complexes to a carrier composed of insoluble finely divided particles,

(b) at least one said second individual container containing a reagent consisting essentially of a buffer solution for diluting or dissolving the sample and for maintaining a reaction mixture of said sample and said suspension at a suitable pH and ionic strength for said agglutination reaction, and

(c) at least one said third individual container containing a reagent consisting essentially of a fixing compound in a concentration sufficient for fixing agglutinates formed from the reaction between the substance to be measured and the sensitized carrier particles, wherein said kit is used for measuring the quantity or concentration of the substance to be measured in the sample by

(i) mixing an amount effective for said measurement of said sensitized carrier particles with said sample dissolved or suitably diluted with a pH stabilizing amount of said buffer solution,

(ii) adding the fixing compound in an amount effective to fix the agglutinates formed in step (i), and

(iii) measuring the absorbancy or turbidity of the reaction mixture from the above step (ii) to determine the quantity or concentration of the substance to be measured in the sample.

2. The measuring reagent kit of claim 1 wherein the insoluble finely divided carrier particles are selected from the group consisting of polystyrene latex, carboxylated polystyrene latex and styrene-divinylbenzene copolymer latex.

3. The measuring reagent kit of claim 1, wherein said fixing compound is selected from the group consisting of 1-ethyl-3-(3-dimethyl-aminopropyl)carbodiimide, 1-cyclohexyl-3-(2-morpholinoethyl)carbodiimide, N-ethyl-5-phenylisoxazonium-3-sulfonate, succinyl chloride, tannic acid, perchloric acid, trichloroacetic acid, thiocyanic acid, and thioglycolic acid.

4. A measuring reagent kit for an immuno-chemical agglutination inhibition reaction measurement of a quantity or concentration of a substance to be measured selected from the group consisting of an antigen, an antibody, and an antigen-antibody complex in a sample, said kit comprising:

a mercantile container, said mercantile container containing at least first, second, third and fourth individual containers;

(a) at least one said first individual container containing a reagent consisting essentially of a suspension of sensitized carrier particles formed by binding at least one substance selected from the group consisting of antigens, antibodies, and antigen-antibody complexes to a carrier composed of insoluble finely divided particles,

(b) at least one said second individual container containing a reagent consisting essentially of a buffer solution for diluting or dissolving the sample and for maintaining a reaction mixture of said sample and said suspension at a suitable pH and ionic strength for said agglutination reaction,

(c) at least one said third individual container containing a reagent consisting essentially of an agglutination agent for inducing said agglutination by reaction with the sensitized carrier particles in the absence of the substance to be measured, and

(d) at least one said fourth individual container containing a reagent consisting essentially of a fixing compound in a concentration sufficient for fixing agglutinates formed from the reaction between the agglutinating agent and the sensitized carrier particles,

wherein said kit is used for measuring the quantity or concentration of the substance to be measured in the sample by

(i) mixing an amount effective for said measurement of said sensitized carrier particles and the agglutination agent with a sample dissolved or suitably diluted with a pH stabilizing amount of said buffer solution,

(ii) adding the fixing compound to the above mixture in an amount effective to fix the agglutinates formed in (i) above, and

(iii) measuring the absorbancy or turbidity of the reaction mixture from the above step (ii) to determine the quantity or concentration of the substance to be measured in the sample.

5. The measuring reagent kit of claim 4 wherein the insoluble finely divided carrier particles are selected from the group consisting of polystyrene latex, carboxylated polystyrene latex and styrene-divinylbenzene copolymer latex.

6. The measuring reagent kit of claim 4 wherein the agglutinating agent is selected from the group consisting of an antigen, antibody and antigen-antibody complex either unbound or bound to a carrier composed of insoluble finely divided particles.

7. The measuring reagent kit of claim 4, wherein said fixing compound is selected from the group consisting of 1-ethyl-3-(3-dimethyl-aminopropyl)carbodiimide, 1-cyclohexyl-3-(2-morpholinoethyl)carbodiimide, N-ethyl-5-phenylisoxazonium-3-sulfonate, succinyl chloride, tannic acid, perchloric acid, trichloroacetic acid, thiocyanic acid, and thioglycolic acid.