Automated immunoassay

• US 4,128,628 A
• Filed: 02/03/1977
• Issued: 12/05/1978
• Est. Priority Date: 03/12/1976
• Status: Expired due to Term

• Current Assignee: University of Virginia Patent Foundation (University of Virginia)
• Original Assignee: The University of Virginia Patent Foundation
• Inventors: Price, Michael G., Terasaki, Wesley L., Brooker, Gary L.
• Primary Examiner(s): Padgett, Benjamin R.
• Assistant Examiner(s): Nucker, Christine M.
• Application Number:
  US05/765,348
• Time in Patent Office: 670 Days
• Field of Search: 424/1, 424/1.5, 424/12, 23/230 B, 23/253 R, 195/103.5 R
• US Class Current: 436/542
• CPC Class Codes: G01N 33/5302 Apparatus specially adapted for immunological test procedures
• CPC Class Codes: G01N 33/531 Production of immunochemical test materials
• CPC Class Codes: G01N 33/538 by sorbent column, particles or resin strip , i.e. sorbent materials
• CPC Class Codes: Y10S 435/973 Simultaneous determination of more than one analyte
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/808 Automated or kit
• CPC Class Codes: Y10S 436/817 Steroids or hormones

Abstract

An automated immunoassay apparatus which provides: time controlled incubation of a multiplicity of sample solutions, each of which comprise a mixture of (a) a sample, possibly containing an antigen or antibody intended to be assayed; (b) a solution of a known concentration of a tagged antigen or tagged antibody; and when assaying for antigen, optionally; (c) a solution of a known titer of an antibody reactable with said antigen;

Isolation of incubated samples from additional sample solutions introduced into said apparatus for assay, whereby the system is isolated at a first predetermined signal from a timing device, and whereby the system is reconnected at a second predetermined signal from the timing device, wherein the second predetermined time period is set so as to enable sufficient time to complete a measurement of the amount of tagged material before the next incubated sample arrives in the detector for the next sequential measurement.

Claims

1. An automated immunoassay apparatus which comprises:

means for time controlled incubation of a multiplicity of sample solutions, each of which comprises a mixture of (a) a sample, possibly containing an antigen or antibody intended to be assayed, (b) a solution of a known concentration of a tagged antigen or tagged antibody, and when assaying for antigen, optionally (c) a solution of a known titer of an antibody reactable with said antigen;

activating detector means for determining the presence of the incubated sample and for activating a timing means upon detection of said sample at a detection station;

means for introducing said incubated sample into said activating detector means;

timing means, which is activatable by said activating detector means when said detector means detects the presence of the incubated sample at a detection station, for generating control signals at predetermined time sequences;

means for separating said incubated sample into a first portion containing antigen or antibody which has not been complexed during the said incubation, and a second portion containing antigen or antibody which has been complexed during said incubation;

means for introducing one of said portions from said separating means into a measuring detector means;

measuring detector means for detecting the amount of tagged material in said portion while said portion is maintained in a virtually static condition;

recording means coupled to said detector means for recording the amounts of tagged material in said measuring detector means;

means for isolating said incubated sample from additional sample solutions introduced into said apparatus for assay, wherein said isolating means is activatable by a control signal emitted from said timing means, whereby said isolating means isolates the system at a first predetermined signal from said timing means, and whereby said isolating means reconnects the system to said incubation means at a second predetermined signal from said timing means, wherein the second predetermined time period is set so as to enable sufficient time to complete said measurement before the next incubated sample arrives in the said detector means for the next sequential measurement; and

means for flushing said portion from said system after said measurement thereof so as to prepare said detector for the next sequential sample for said measurement.

2. The apparatus of claim 1 wherein said activating detector means is a conductivity detector for detecting changes in conductivity as a fluid is passed therethrough.

3. The apparatus of claim 1, wherein said activating detector means is a photodetection which is capable of detecting changes in solution turbidity.

4. The apparatus of claim 1, wherein said activating detector means is a colorimeter which is capable of determining the presence of dyes in said solution.

5. The apparatus of claim 1, wherein said activating detector means is a pH meter which is capable of detecting changes in solution pH.

6. The apparatus of claim 1, wherein said activating detector means is a refractometer which is capable of detecting changes in solids content of the solution as it is passed therethrough.

7. The apparatus of claim 1, wherein said measuring detector means is a colorimeter which is capable of detecting changes in solution color due to the presence of dye.

8. The apparatus of claim 1 wherein said measuring detector means is an enzyme substrate which is specific for a particular enzyme tagging material.

9. The apparatus of claim 1, wherein said measuring detector means is a radioactivity detector.

10. A method for continuous immunoassay, which comprises:

incubating for a predetermined time period, a multiplicity of sample solutions, each of which comprises a mixture of (a) a sample possibly containing an antigen intended to be assayed;

(b) a solution of a known concentration of a tagged antigen; and

when assaying for antigen, optionally, (c) a solution containing a known titer of antibody reactable with said antigen, the concentration of said antibody being selected so as to form an incubated sample containing an antibody-antigen complex, and possibly containing unreacted antigen and tagged antigen, if said sample contains an antigen;

directing said incubated sample into an activating detector which is capable of determining the presence of said sample and activating,

activating a timing means upon detection of said sample in said detector;

isolating said incubated sample from any additional samples being incubated on a predetermined signal from said timing means;

separating said incubated solution into a first portion containing an antigen or antibody which has not been complexed during the said incubation and a second portion containing antigen or antibody which has been complexed during said incubation;

directing one of said portions into a measuring detector, detecting the amount of tagged material in said portion while said portion is maintained in a virtually static condition, and recording said measurement;

evacuating said portion to waste;

directing a subsequent incubated sample into said activating detector on a second predetermined signal from said timing means so that the next subsequent incubated sample to be assayed arrives in the activating detector for the next measurement after the previous measurement is completed; and

comparing the measurement with precalibrated values to determine the quantity of antigen being assayed.

11. The method of claim 10, wherein said samples contain antibodies intended to be assayed.

12. The method of claim 11, wherein said mixture contains a sample containing antibodies intended to be assayed and a solution of a known concentration of a tagged antigen.

13. The method of claim 10, wherein the presence of sample in said activating detector means is detected by change in pH or conductivity of said solution.

14. The method of claim 10, wherein the presence of sample in said activating detector means is detected by change in refractive index of said solution.

15. The method of claim 13, wherein the tagged antigen or antibody is tagged with a radioactive isotope and wherein measurement by said measuring means is effected by detection of quantity of radioactivity as a function of time.

16. The method of claim 10, wherein the tagged antigen or antibody is tagged with a dye and wherein measurement by said measuring means is effected by detection of color.