Method of and apparatus for immune analysis

• US 5,750,410 A
• Filed: 01/26/1996
• Issued: 05/12/1998
• Est. Priority Date: 08/26/1994
• Status: Expired due to Term

• Current Assignee: Kyoto Daiichi Kagaku Company Limited
• Original Assignee: Kyoto Daiichi Kagaku Company Limited
• Inventors: Yamaguchi, Yoshinori, Dou, Xiaoming, Uenoyama, Harumi
• Primary Examiner(s): Spiegel, Carol A.
• Application Number:
  US08/592,522
• Time in Patent Office: 837 Days
• Field of Search: 422/82.05, 422/82.09, 422/82.11, 435/962, 436/525, 436/164, 436/524, 436/536
• US Class Current: 436/525
• CPC Class Codes: G01N 21/552 Attenuated total reflection
• CPC Class Codes: G01N 21/77 by observing the effect on a chemical indicator
• CPC Class Codes: G01N 33/585 with a particulate label, e.g. coloured latex
• CPC Class Codes: Y10S 435/962 Prevention or removal of interfering materials or reactants or other treatment to enhance results, e.g. determining or preventing nonspecific binding

Abstract

A total reflection cell has a total reflection prism on at least one surface thereof. A mixed solution containing a gold colloid labelled antibody which is adsorbed on a gold colloid is stored in the total reflection cell, and a sample solution containing an antigen causing antigen-antibody reaction with the antibody is added thereto for forming a gold colloid labelled immune complex. A measuring beam is introduced into the total reflection prism from an incident optical system at an angle θ of incidence causing total reflection and an outgoing beam from the total reflection prism is received by a measuring optical system, thereby measuring absorption by the gold colloid labelled immune complex and carrying out qualification and determination of the antigen.

Claims

1. A method for quantitatively determining a target antigen or a target antibody in a liquid sample in an immunoassay, comprising:

(a) reacting said liquid sample with a solution comprising an antibody or antigen labelled with a colloidal noble metal particle, wherein said antibody or antigen specifically binds to said target antigen or said target antibody, respectively, to form a liquid sample/solution mixture containing a labelled immune complex;

(b) contacting said liquid sample/solution mixture with a total reflection cell having a surface comprising a total reflection prism;

(c) illuminating said total reflection prism with a measuring beam of light having a wavelength in a range of visible to infrared light at an angle of incidence which causes total reflection at an interface between said total reflection prism and said liquid sample/solution mixture in said total reflection cell;

(d) measuring absorption of said measuring beam at the interface; and

(e) comparing said measured absorption to a standard absorption obtained from a known concentration of said target antigen or said target antibody processed according to steps (a)-(d) to determine the amount of said target antigen or said target antibody in said liquid sample.

2. The method in accordance with claim 1, wherein said reacting step (a) is carried out in said total reflection cell.

3. The method in accordance with claim 1, wherein said reacting step (a) is carried out in a reaction container, and said liquid sample/solution mixture is thereafter transferred into said total reflection cell in said contacting step (b).

4. The method in accordance with claim 1, wherein said colloidal noble metal particle label is a colloidal gold, silver or copper particle label.

5. A method for qualitatively determining a target antigen or a target antibody in a liquid sample in an immunoassay, comprising:

(a) reacting said liquid sample with a solution comprising an antibody or antigen labelled with a colloidal noble metal particle, wherein said antibody or antigen specifically binds to said target antigen or said target antibody, respectively, to form a liquid sample/solution mixture containing a labelled immune complex;

(b) contacting said liquid sample/solution mixture with a total reflection cell having a surface comprising a total reflection prism;

(c) illuminating said total reflection prism with a measuring beam of light having a wavelength in a range of visible to infrared light at an angle of incidence which causes total reflection at an interface between said total reflection prism and said liquid sample/solution mixture in said total reflection cell; and

(d) measuring absorption of said measuring beam at the interface to determine the presence of said target antigen or said target antibody in said liquid sample.