Apparatus and process for simplified measurement

• US 5,552,276 A
• Filed: 03/09/1994
• Issued: 09/03/1996
• Est. Priority Date: 03/18/1993
• Status: Expired due to Fees

• Current Assignee: Mochida Pharmaceutical Company Limited
• Original Assignee: Mochida Pharmaceutical Company Limited
• Inventors: Miyauchi, Yasuhiko, Katamine, Tomoaki, Matsuura, Takashi, Mochida, Ei
• Primary Examiner(s): Housel, James C.
• Assistant Examiner(s): CHIN, CHRISTOPHER L
• Application Number:
  US08/207,715
• Time in Patent Office: 909 Days
• Field of Search: 422/55-58, 422/61, 422/101, 435/6, 435/7.4, 435/7.5, 435/7.9, 435/7.92, 435/7.93, 435/7.94, 435/7.1, 435/174, 435/176, 435/177, 435/179-181, 435/287-289, 435/291, 435/805, 435/810 , 435/970, 435/973, 436/514, 436/518, 436/524, 436/528, 436/530, 436/164, 436/169, 436/170, 436/805, 436/807, 436/810, 436/824, 436/827, 436/525, 436/531, 436/532-534, 436/172, 436/800 , 436/804, 436/527
• US Class Current: 435/6.19
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick

Abstract

A simplified measuring apparatus for use in the qualitative or quantitative measurement of substances to be assayed in test samples, such as proteins, antibodies and the like, in a small amount of test samples by a simple and easy operation without requiring a B/F separation step comprises: (a) a liquid permeable porous reaction membrane whose surface having at least one reaction area to which an affinity substance capable of directly or indirectly capturing a substance to be assayed or a soluble agent is immobilized; (b) a porous body arranged on the upper part of the porous reaction membrane, to which a soluble agent capable of being solubilized by the addition of a test sample is adhered in a releasable manner; (c) an absorption member arranged on the lower part of the porous reaction membrane, which contacts with a periphery, excluding the reaction area, of the porous reaction membrane via a liquid non-permeable sheet; (d) a liquid non-permeable transparent cover arranged on the lower part of the absorption member; and (e) a liquid non-permeable case which encloses the foregoing members (a) to (d), having an opening for use in test sample introduction on its top surface and another opening for observation on its under surface.

Claims

1. A simplified measuring apparatus for use in a binding assay to determine the presence or amount of an analyte in a fluid test sample through the use of a label capable of producing a detectable response, said apparatus comprising:

(a) a porous body having releasably attached thereto an agent soluble in said test sample;

(b) a liquid permeable porous reaction membrane disposed below said porous body having defined thereon at least one reaction area, said reaction area having immobilized thereon an affinity substance capable of directly or indirectly capturing the analyte or an agent soluble in said test sample to thereby produce the detectable response;

(c) an absorption member disposed below said liquid permeable porous membrane having an opening being positioned below the reaction area of said liquid permeable porous membrane to define a reaction solution storing space within the opening, said absorption member being, arranged so as to contact only a peripheral area of said liquid permeable porous membrane via an intervening liquid impermeable sheet;

(d) a liquid impermeable transparent cover disposed below said opening of said absorption member; and

(e) a liquid impermeable case accommodating said members (a), (b), (c) and (d) having a top surface adjacent said porous body and a bottom surface adjacent said transparent cover, said top surface having defined therein a top opening for introducing said fluid test sample, and said bottom surface having defined therein a bottom opening for observation of said detectable response.

2. A simplified measuring apparatus according to claim 1, wherein said reaction area comprises a plurality of reaction sites each having immobilized thereon an affinity substance different from other reaction sites.

3. A simplified measuring apparatus according to claim 2, wherein said plurality of reaction sites are separately defined from each other.

4. A simplified measuring apparatus according to claim 1, wherein said affinity substance immobilized on said liquid permeable porous reaction membrane is a member selected from the group consisting of an antibody and an antigen.

5. A simplified measuring apparatus according to claim 1, wherein said affinity substance immobilized on said liquid permeable porous reaction membrane is a member selected from the group consisting of a nucleic acid, a lectin, biotin, avidin, an enzyme, an inhibitor and a receptor.

6. A simplified measuring apparatus according to claim 1, wherein said agent soluble in said test sample is a member selected from the group consisting of an antibody and an antigen, that has been labeled with the label to produce the detectable response.

7. A simplified measuring apparatus according to claim 6, wherein said label is a member selected from the group consisting of an enzyme, a fluorescent compound, a radioactive label, a chemiluminescent compound, a colloidal metal particle, and a nonmetal particle.

8. A simplified measuring apparatus according to claim 7, wherein said label is a nonmetal particle selected from the group consisting of a dye particle, and a latex particle.

9. A simplified measuring apparatus according to claim 1, wherein said agent soluble in said test sample is a member selected from the group consisting of a nucleic acid, a lectin, biotin, avidin, an enzyme, an inhibitor and a receptor, that has been labeled with the label to produce the detectable response.

10. A simplified measuring apparatus according to claim 9, wherein said label is a member selected from the group consisting of an enzyme, a fluorescent compound, a radioactive label, a chemiluminescent compound, a colloidal metal particle, and a nonmetal particle.

11. A simplified measuring apparatus according to claim 10, wherein said label is a nonmetal particle selected from the group consisting of a dye particle, and a latex particle.

12. A simplified measuring apparatus according to claim 1, wherein said liquid permeable porous reaction membrane is a member selected from the group consisting of cellulose, a cellulose derivative, a porous synthetic polymer, a glass fiber filter, and a fabric.

13. A simplified measuring apparatus according to claim 12, wherein said liquid permeable porous reaction membrane is a cellulose derivative consisting of nitrocellulose.

14. A simplified measuring apparatus according to claim 1, wherein said porous body is a member selected from the group consisting of cellulose, a cellulose derivative, a porous synthetic polymer, a glass fiber filter, and a fabric.

15. A simplified measuring apparatus according to claim 1, wherein said absorption member is a member selected from the group consisting of cellulose, a cellulose derivative, a porous synthetic polymer, a glass fiber filter, a nonwoven fabric, a granular absorbent, and a fabric.

16. A simplified measuring apparatus according to claim 1, wherein said liquid permeable porous reaction membrane is nitrocellulose, and said absorption member and said porous body comprise cellulose.

17. A process for determining the presence or quantity of an analyte in a fluid test sample through the use of the simplified measuring apparatus of claim 1, comprising the steps of

(a) adding the fluid test sample that is suspected to contain the analyte to the top opening defined on the top surface of the case to allow dissolution of said soluble agent releasably attached to said porous body and subsequent migration of the released agent to said porous liquid permeable reaction membrane;

(b) allowing said analyte, said released agent, or a complex thereof to contact with and bind to said affinity substance immobilized on the reaction area of said liquid permeable porous reaction membrane to thereby directly or indirectly produce the detectable response;

(c) allowing the fluid test sample added and said released agent that failed to become immobilized onto said liquid permeable porous reaction membrane to pass through said liquid permeable porous reaction membrane for absorption by said absorption member; and

(d) observing said detectable response from said bottom opening on said bottom surface of the case to determine the presence or amount of the analyte in the fluid test sample.

18. A process according to claim 17, wherein said reaction area in said liquid permeable porous reaction membrane is washed to remove unbound materials.

19. A process according to claim 17, wherein said fluid test sample is a member selected from the group consisting of a body fluid, and a solution obtained by one of dissolving and extraction-diluting a biological component.