Specific coupling reaction measuring method

• US 7,553,676 B2
• Filed: 08/21/2006
• Issued: 06/30/2009
• Est. Priority Date: 11/12/2002
• Status: Active Grant

• Current Assignee: PHC Holdings Corporation
• Original Assignee: Panasonic Corporation (Panasonic Holdings Corporation)
• Inventors: Kawamura, Tatsurou, Shigetoh, Nobuyuki, Kamei, Akihito
• Primary Examiner(s): Nguyen; Bao-Thuy L
• Assistant Examiner(s): BROWN, MELANIE YU
• Application Number:
  US11/507,011
• Publication Number:
  US 20060281195A1
• Time in Patent Office: 1,044 Days
• Field of Search: 436/518, 436/524, 436/525, 436/526, 436/164, 435/7.1, 435/287.2
• US Class Current: 436/526
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/54333 Modification of conditions of immunological binding reaction, e.g. use of more than one type of particle, use of chemical agents to improve binding, choice of incubation time or application of magnetic field during binding reaction

Abstract

A specific coupling reaction measuring method of this invention for measuring a content of a subject substance in a sample includes the steps of (a) constructing a reaction system including the sample and a magnetic particle on which a specific coupling substance for specifically coupling with the subject substance is immobilized; (b) measuring an optical characteristic of the reaction system; and (c) removing, from the reaction system, an agglutination complex including the subject substance, the specific coupling substance and the magnetic particle by utilizing magnetic force.

Claims

1. A specific coupling reaction measuring method for measuring a plurality of subject substances in a sample, comprising the steps of:

(a) constructing a reaction system including said sample and a specific coupling substance for specifically coupling with a first subject substance;

(b) measuring an optical characteristic of said reaction system;

(c) adding, to said reaction system, a magnetic complex including a magnetic particle and a substance capable of coupling with an agglutination complex including said first subject substance and said specific coupling substance;

(d) removing, by utilizing magnetic force, said agglutination complex coupled with said magnetic complex from said reaction system;

(e) adding, to said reaction system, another specific coupling substance for specifically coupling with another subject substance; and

(f) measuring an optical characteristic of said reaction system after the step (e).

2. The specific coupling reaction measuring method of claim 1,

wherein said optical characteristic is an intensity of scattered light or a quantity of transmitted light.

3. The specific coupling reaction measuring method of claim 1,

wherein each said magnetic particles has a diameter of approximately 0.05 through 2 μ

m.

4. The specific coupling reaction measuring method of claim 1,

wherein a portion of said magnetic particles are included in said agglutination complex and other magnetic particles remain in said reaction system in the step (a), and said other magnetic particles remaining in said reaction system are removed from said reaction system in the step (d).

5. The specific coupling reaction measuring method of claim 1, wherein said reaction system includes 2 through 6 wt % of polyethylene glycol in the step (e).

6. The specific coupling reaction measuring method of claim 1, wherein said another specific coupling substance is composed of an antigen or antibody for specifically coupling with said another subject substance and a nonmagnetic particle on which said antigen or antibody is immobilized.

7. The specific coupling reaction measuring method of claim 6, wherein said magnetic particle has a diameter of approximately 0.05 through 2 μ

m and said nonmagnetic particle has a diameter of approximately 0.05 through 2 μ

m.

8. The specific coupling reaction measuring method of claim 1, wherein a combination of said first subject substance and said another subject substance is a combination of human hemoglobin and human albumin.