EP0103426A2
Expired / Lapsed
EP
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Immunoassay apparatus and method
Assignee: Individual
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Filing: 1983-08-22
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Granted: —
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Expires: 2003-08-22
Abstract
Abstract Fluorescent immunoassay apparatus and method employing a disposable consisting, in a preferred embodiment, of a short length of precise diameter capillary tubing having an axially disposed optical fiber to which is immobilized a monolayer of a component of the antibody-antigen complex (e.g., an antibodyl, an inert diluent, and a preload of a known amount of tagged complement to the immobilized component (e.g., a fluorescent-tagged antigen). The dimensions of the fiber and the capillary tubing are chosen so as to allow the tube to fill itself by capillary action once an end of the tube is immersed into the sample. Precise timing and ballistic measurements may, if desired, be avoided by insuring the incubation time is larger than the diffusion time necessary to scavenge the volume between the fiber and the capillary tubing. Fluorometric measurement is made by total reflection fluorescence techniques.
Inventors
- 👤 Tomas B. Hirschfeld
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-28 22:53
CPC Classifications
G; G01; G01N; G01N21/00; G01N21/62; G01N21/63; G01N21/64; G01N21/645; G01N21/648; G01N21/6428; G01N21/75; G01N21/77; G01N21/7703; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54366; G01N33/54373; G01N21/01; G01N21/03; G01N2021/0346; Y; Y10; Y10S; Y10S436/00; Y10S436/805; Y10S436/807
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
112 words
An apparatus for performing immunoassays using an antigen-antibody complex incorporating a fluorescent tag capable of emitting fluorescent radiation when excited by exciting radiation, characterized in that it comprises an optical fiber (12) transmissive to both the exciting radiation and the fluorescent radiation, a coating (32) on at least a portion (30) of the fiber (12), the coating (32) having a plurality of sites (44) each being capable of having attached thereto a selected moiety (46) of the antigen-antibody complex so as to leave substantially unaffected the activity of the selected moiety for forming the antigen-antibody complex and means for controlling a volume of capillary dimensions bounded by the portion of the fiber.
Claim 2
Independent Claim
112 words
An apparatus for performing immunoassays using an antigen-antibody complex incorporating a fluorescent tag capable of emitting fluorescent radiation when excited by exciting radiation, characterized in that it comprises an optical fiber (12) transmissive to both the exciting radiation and the fluorescent radiation, a coating (32) on at least a portion (30) of the fiber (12), the coating (32) having a plurality of sites (44) each being capable of having attached thereto a selected moiety (46) of the antigen-antibody complex so as to leave substantially unaffected the activity of the selected moiety for forming the antigen-antibody complex and means for controlling a volume of capillary dimensions bounded by the portion of the fiber.
Claim 3
Dependent Claim
35 words
An apparatus according to claim 1, characterized in that the coating is on a predetermined length (30) of the fiber (12), a plurality of selected moieties of the antigen-antibody complex being attached to the coating.
Claim 4
Dependent Claim
22 words
An apparatus according to claim 2, characterized in that the selected moieties are attached to substantially all of the plurality of sites.
Claim 5
Dependent Claim
53 words
An apparatus according to claim 2, characterized in that the selected moieties (46) and a plurality of particles of an immunologically inert material (56) are attached in a predetermined ratio to substantially all of the plurality of sites (44), a plurality of tagged complement (50) to the moieties being complexed to the moieties.
Claim 6
Dependent Claim
49 words
An apparatus according to claim 1, characterized in that the capillary dimensions are predetermined, the means for controlling comprising an elongate enclosure means dimensioned to surround the fiber in a spaced apart relationship of capillary dimensions and means for positioning the fiber in the relationship within the enclosure means.
Claim 7
Dependent Claim
34 words
An apparatus according to claim 5, characterized in that the elongate enclosure means is a capillary tube (14, 114), at least a portion of the interior thereof being provided a reagent coating (30, 130)
Claim 8
Dependent Claim
63 words
An apparatus according to claim 6, characterized in that the means for positioning the fiber includes a substantially centrally apertured stopper (16) mounted in a first end of the capillary tube (14), the fiber (12) being positioned in an aperture (22) thereof, the stopper (16) being provided with locating means (21) for defining the location of an end (24) of the fiber (12).
Claim 9
Dependent Claim
32 words
An apparatus according to claim 7, characterized in that the opposite end of the capillary tube (14) is dimensioned to have a smaller inside diameter than the remainder of the capillary tube.
Claim 10
Dependent Claim
98 words
An apparatus according to claim 1, characterized in that a fluorescent tag (50, 52) capable of emitting fluorescent radiation when excited by more energetic exciting radiation is incorporated in a constituent of an antigen-antibody complex (46), the capillary tube (14) being dimensioned to accept the fiber (12) in spaced-apart relationship and a stopper being affixed to a first end of the capillary tube, for supporting the fiber substantially coaxially within the capillary tube with the fiber passing through the stopper substantially centrally, the stopper having a locating means for defining the location of an end of the fiber.
Claim 11
Independent Claim
202 words
A method of immunoassay employing an optical element transmissive to excitation radiation which is adapted to excite fluorescence of a fluorescent tag included as a constituent of an antigen-antibody complex the element being also transmissive to the fluorescence, the element having selected moieties of the antibody-antigen complex attached to a predetermined area of its surface, the method being characterized in that the optical element is immersed in a fluid sample to be assayed so that the portion of the sample immunologically reactive with the selected moieties can react therewith to form the complex, the immersion of the optical element being controlled in the fluid sample so as to ensure that the predetermined area is completely covered with a layer of known thickness of the fluid sample, incubating said optical element in said layer of fluid sample for a time in excess of the time required for diffusional processes to scavenge the volume of sample defined by the known area and the known thickness, the optical element being illuminated with an evanescent electromagnetic wave of the excitation radiation so as to excite fluorescence of the tag in the complex attached to the optical element and the intensity o.f the fluorescence being then measured.
Claim 12
Independent Claim
202 words
A method of immunoassay employing an optical element transmissive to excitation radiation which is adapted to excite fluorescence of a fluorescent tag included as a constituent of an antigen-antibody complex the element being also transmissive to the fluorescence, the element having selected moieties of the antibody-antigen complex attached to a predetermined area of its surface, the method being characterized in that the optical element is immersed in a fluid sample to be assayed so that the portion of the sample immunologically reactive with the selected moieties can react therewith to form the complex, the immersion of the optical element being controlled in the fluid sample so as to ensure that the predetermined area is completely covered with a layer of known thickness of the fluid sample, incubating said optical element in said layer of fluid sample for a time in excess of the time required for diffusional processes to scavenge the volume of sample defined by the known area and the known thickness, the optical element being illuminated with an evanescent electromagnetic wave of the excitation radiation so as to excite fluorescence of the tag in the complex attached to the optical element and the intensity o.f the fluorescence being then measured.
References Cited (Backward Citations)
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