Fluorimetric immunological assay with magnetic particles

• US 4,731,337 A
• Filed: 07/24/1985
• Issued: 03/15/1988
• Est. Priority Date: 07/26/1984
• Status: Expired due to Term

• Current Assignee: Labsystems OY Pulttitie 9
• Original Assignee: Labsystems Oy (Thermo Fisher Scientific Incorporated)
• Inventors: Luotola, Juhani E. I., Harjunmaa, Hannu
• Primary Examiner(s): NOT, DEFINED
• Assistant Examiner(s): NOT, DEFINED
• Application Number:
  US06/758,539
• Time in Patent Office: 965 Days
• Field of Search: 436/526, 436/533, 436/538, 436/806, 436/824
• US Class Current: 436/526
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y10S 436/806 Electrical property or magnetic property
• CPC Class Codes: Y10S 436/824 Immunological separation techniques

Abstract

Fluorometric immunological assay method in which an antigen is attached onto magnetic particles, which are, for the time of measurement, pulled against the wall of the measurement vessel by means of the magnetic field. The magnetic particles adhering to the vessel walls are irradiated and the resultant emitted fluorescence is measured.

Claims

1. A method of immunological assay for a target antibody or target antigen comprising:

a. attaching an antigen or antibody to solid magnetic particles;

b. contacting the magnetic particles resulting from (a) with

(1) a sample containing either the target antibody or target antigen, and

(2) fluorescently labeled antibody or fluorescently labeled antigen, respectively, said contacting taking place in a measurement vessel;

c. forcing the magnetic particles resulting from (b) against the wall of the measuring vessel by an applied magnetic field;

d. irradiating, through the walls of the measuring vessel, the magnetic particles adhering to the wall as a result of (c); and

e. measuring the resultant fluorescent or phosphorescent radiation with a detector after this radiation has passed through the walls of the measuring vessel.

2. Method as claimed in claim 1, characterized in that the solid magnetic particles to which the antigen or antibody is attached comprises polymer particles to which a magnetic substance has been added.

3. Method as claimed in claim 1, characterized in that, before measurement, a colouring agent is added in the measurement vessel, said colouring agent having a strong absorption at the wavelength of the exitation radiation.

4. Method as claimed in claim 1, characterized in that, before measurement, a colouring agent is added into the measurement vessel, said colouring agent having a strong absorption at the wavelength of the emission radiation.

5. Method as claimed in claim 3, characterized in that the colouring agent is black.

6. Method as claimed in claim 1, characterized in that both the excitation radiation is passed into the sample and the fluorescence or phosphorescence radiation is collected to the detector through the bottom of the measurement vessel.

7. Method as claimed in claim 2, characterized in that, before measurement, a colouring agent is added into the measurement vessel, said colouring agent having a strong absorption at the wavelength of the excitation radiation.

8. Method as claimed in claim 2, characterized in that, before measurement, a colouring agent is added into the measurement vessel, said colouring agent having a strong absorption at the wavelength of the emission radiation.

9. Method as claimed in claim 3, characterized in that, before measurement, a colouring agent is added into the measurement vessel, said colouring agent having a strong absorption at the wavelength of the emission radiation.

10. Method as claimed in claim 7, characterized in that, before measurement, a colouring agent is added into the measurement vessel, said colouring agent having a strong absorption at the wavelength of the emission radiation.

11. Method as claimed in claim 4, characterized in that the colouring agent is black.

12. Method as claimed in claim 7, characterized in that the colouring agent is black.

13. Method as claimed in claim 8, characterized in that the colouring agent is black.

14. Method as claimed in claim 9, characterized in that the colouring agent is black.

15. Method as claimed in claim 10, characterized in that the colouring agent is black.

16. Method as claimed in claim 2, characterized in that both the excitation radiation is passed into the sample and the fluorescence or phosphorescence radiation is collected to the detector through the bottom of the measurement vessel.

17. Method as claimed in claim 3, characterized in that both the excitation radiation is passed into the sample and the fluorescence or phosphorescence radiation is collected to the detector through the bottom of the measurement vessel.

18. Method as claimed in claim 4, characterized in that both the excitation radiation is passed into the sample and the fluorescence or phosphorescence radiation is collected to the detector through the bottom of the measurement vessel.

19. Method as claimed in claim 5, characterized in that both the excitation radiation is passed into the sample and the fluorescence or phosphorescence radiation is collected to the detector through the bottom of the measurement vessel.

20. Method as claimed in claim 7, characterized in that both the excitation radiation is passed into the sample and the fluorescence or phosphorescence radiation is collected to the detector through the bottom of the measurement vessel.

21. Method as claimed in claim 8, characterized in that both the excitation radiation is passed into the sample and the fluorescence or phosphorescence radiation is collected to the detector through the bottom of the measurement vessel.

22. Method as claimed in claim 9, characterized in that both the excitation radiation is passed into the sample and the fluorescence or phosphorescence radiation is collected to the detector through the bottom of the measurement vessel.

23. Method as claimed in claim 10, characterized in that both the excitation radiation is passed into the sample and the fluorescence or phosphorescence radiation is collected to the detector through the bottom of the measurement vessel.

24. Method as claimed in claim 11, characterized in that both the excitation radiation is passed into the sample and the fluorescence or phosphorescence radiation is collected to the detector through the bottom of the measurement vessel.

25. Method as claimed in claim 12, characterized in that both the excitation radiation is passed into the sample and the fluorescence or phosphorescence radiation is collected to the detector through the bottom of the measurement vessel.

26. Method as claimed in claim 13, characterized in that both the excitation radiation is passed into the sample and the fluorescence or phosphorescence radiation is collected to the detector through the bottom of the measurement vessel.

27. Method as claimed in claim 14, characterized in that both the excitation radiation is passed into the sample and the fluorescence or phosphorescence radiation is collected to the detector through the bottom of the measurement vessel.

28. Method as claimed in claim 15, characterized in that both the excitation radiation is passed into the sample and the fluorescence or phosphorescence radiation is collected to the detector through the bottom of the measurement vessel.