US4477578A
Expired / Lapsed
US
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Method and apparatus for performing assays
Assignee: Vitek Systems Inc
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Filing: 1982-03-04
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Granted: 1984-10-16
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Expires: 2002-03-04
Abstract
Abstract Method and apparatus are provided for carrying out multiple simultaneous transfers of fluid. The method and apparatus are particularly directed toward immunoassays wherein immunologically active compounds, such as antigens and haptens, are detected through their associated antibodies. The device relies on the ability to transfer fluids, such as biological samples and reagents, between a reservoir and an associated receptacle. By providing a receptacle having a port at its lower end and which is otherwise hermetically sealed, such fluid transfer can be effected by immersing the port beneath the surface of the fluid in the reservoir and manipulating the pressure on the remaining surface area outside the port. The transfer of biological fluids at positive pressure provides enhanced fluids flow characteristics, particularly reduction or elimination of the tendency of these fluids to froth or bubble. Moreover, since the fluids can easily be manipulated, they can be agitated to speed up the reaction and reduce the overall reaction time and can be transferred from the reaction zone to allow interim measurements of the extent of reaction to provide for a rate mode assay. The method and apparatus also find use in preparing solid phase reagents for use in assay systems, as well as a highly accurate pipetting system in analytic applications not limited to immunoassays.
Inventors
- 👤 Laughton E. Miles
- 👤 Arthur H. Rogers, Jr.
- 👤 Charles H. Rogers
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-31 21:56
CPC Classifications
G; G01; G01N; G01N35/00; G01N35/10; G01N35/1065; G01N1/00; G01N1/28; G01N1/38; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/5302; G01N33/543; G01N33/54366; G01N2035/00465; G01N2035/00534; G01N2035/00544; G01N2035/1027; G01N2035/1048; G01N2035/1055; G01N35/1079; Y; Y10; Y10S; Y10S436/00; Y10S436/807; Y10S436/808; Y10S436/81; Y10S436/82; Y10T; Y10T436/00; Y10T436/11; Y10T436/113332
Claims Analysis
Total Claims: 26
Claim 1
Independent Claim
193 words
A method for determining an analyte which is a member of a specifically binding pair in a sample fluid, said method comprising: introducing into a reaction zone comprising one member of the specifically binding pair immobilized within the interior of a receptacle having a single port (a) the sample fluid by immersing the receptacle port beneath the surface of the sample fluid and elevating the pressure surrounding the receptacle and (b) a labelling reagent comprising a detectable label conjugated to one member of a specifically binding pair by immersing the receptacle port beneath the surface of a solution of said reagent to entrap a fixed amount of gas in the receptacle, and elevating the surrounding pressure to introduce reagent into the receptacle and compress the gas until the pressure inside the receptacle substantially equals the surrounding pressure, whereby a portion of the labelling reagent becomes bound to the reaction zone, the extent of said portion being related to the amount of analyte in the sample fluid; expelling the mixture of said sample fluid and said labelling reagent from the reaction zone by lowering the surrounding pressure; and detecting the amount of bound label.
Claim 2
Independent Claim
193 words
A method for determining an analyte which is a member of a specifically binding pair in a sample fluid, said method comprising: introducing into a reaction zone comprising one member of the specifically binding pair immobilized within the interior of a receptacle having a single port (a) the sample fluid by immersing the receptacle port beneath the surface of the sample fluid and elevating the pressure surrounding the receptacle and (b) a labelling reagent comprising a detectable label conjugated to one member of a specifically binding pair by immersing the receptacle port beneath the surface of a solution of said reagent to entrap a fixed amount of gas in the receptacle, and elevating the surrounding pressure to introduce reagent into the receptacle and compress the gas until the pressure inside the receptacle substantially equals the surrounding pressure, whereby a portion of the labelling reagent becomes bound to the reaction zone, the extent of said portion being related to the amount of analyte in the sample fluid; expelling the mixture of said sample fluid and said labelling reagent from the reaction zone by lowering the surrounding pressure; and detecting the amount of bound label.
Claim 3
Dependent Claim
53 words
A method as in claim 1, wherein the analyte and the substance in the reaction zone are analogous members of one specifically binding pair constituting a ligand and its receptor so that the analyte and the substance compete for binding to the labelled reagent comprising a complementary member of the specifically binding pair.
Claim 4
Dependent Claim
55 words
A method as in claim 1, wherein the analyte and the substance in the reaction zone comprise complementary members of one specifically binding pair so that the analyte is capable of binding directly to the bound substance in competition with the labelled reagent comprising a member of the specifically binding pair analogous to the analyte.
Claim 5
Dependent Claim
44 words
A method as in claim 1, wherein the analyte and the substance are members of different specifically binding pairs, so that the analyte binds to the substance through an intermediate binding substance which is capable of binding to both the analyte and the substance.
Claim 6
Dependent Claim
52 words
A method as in claim 4, wherein the receptacle includes the reaction zone and the intermediate binding substance is introduced into the reaction zone separately from the analyte by immersing the receptacle port beneath the surface of a solution containing said intermediate binding substance and thereafter elevating the pressure in the enclosure.
Claim 7
Independent Claim
37 words
A method as in any of claims 2, 3, 4 or 5, wherein the pressure in the enclosure is periodically varied to agitate the sample fluid in the reaction zone so that the reaction time is reduced.
Claim 8
Independent Claim
34 words
A method as in any of claims 2 or 3 further comprising periodically removing the sample fluid from the reaction zone and determining the extent of binding between the labelled reagent and the substance.
Claim 9
Dependent Claim
32 words
A method as in claim 4, wherein the reservoir includes the reaction zone and the intermediate binding substance is introduced to the reservoir separately from the analyte by means of the receptacle.
Claim 10
Independent Claim
217 words
A method for determining in a sample fluid the presence of an analyte which is a member of a specifically binding pair consisting of a ligand and a receptor, said method comprising: introducing into a receptacle having a single port (a) the sample fluid and (b) a labelled reagent comprising a detectable label conjugated to one member of the specifically binding pair by immersing the receptacle port beneath the surface of the sample fluid and the labelled reagent to entrap a fixed amount of gas therein, and elevating the surrounding pressure by a predetermined amount to introduce said sample fluid and reagent into the receptacle and thereby compress the gas until the pressure inside the receptacle substantially equals the surrounding pressure; expelling the mixture of said sample fluid and said labelled reagent from the receptacle into a reservoir having a reaction zone comprising one member of the specifically binding pair immobilized therein by lowering the surrounding pressure whereby a portion of the labelled reagent becomes bound to the reaction zone, the extent of said portion being related to the amount of analyte in the sample fluid; removing unbound labelled reagent from the reservoir by immersing the port beneath the surface of the fluid and elevating the surrounding pressure; and detecting the amount of the bound labelled reagent.
Claim 11
Independent Claim
217 words
A method for determining in a sample fluid the presence of an analyte which is a member of a specifically binding pair consisting of a ligand and a receptor, said method comprising: introducing into a receptacle having a single port (a) the sample fluid and (b) a labelled reagent comprising a detectable label conjugated to one member of the specifically binding pair by immersing the receptacle port beneath the surface of the sample fluid and the labelled reagent to entrap a fixed amount of gas therein, and elevating the surrounding pressure by a predetermined amount to introduce said sample fluid and reagent into the receptacle and thereby compress the gas until the pressure inside the receptacle substantially equals the surrounding pressure; expelling the mixture of said sample fluid and said labelled reagent from the receptacle into a reservoir having a reaction zone comprising one member of the specifically binding pair immobilized therein by lowering the surrounding pressure whereby a portion of the labelled reagent becomes bound to the reaction zone, the extent of said portion being related to the amount of analyte in the sample fluid; removing unbound labelled reagent from the reservoir by immersing the port beneath the surface of the fluid and elevating the surrounding pressure; and detecting the amount of the bound labelled reagent.
Claim 12
Independent Claim
189 words
A method for determining in a sample fluid the presence of an analyte which is a member of a specifically binding pair, said method comprising: immersing a port in an otherwise closed receptacle beneath the surface of said sample fluid to entrap a fixed amount of gas within the receptacle, said fluid sample being held in a reservoir which is located inside an enclosure; manipulating the pressure within said enclosure to compress said fixed amount of entrapped gas and to transfer the sample fluid and a labelled reagent which is a member of a specifically binding pair between the reservoir and the receptacle to expose a reaction zone comprising an immobilized substance capable of reacting by non-covalent binding with said analyte to said fluid so that the amount of labelled reagent bound within the reaction zone is proportional to the amount of analyte present; and determining the extent of reaction between the analyte and the substance bound within the reaction zone by observing the amount of label bound within the reaction zone after the sample fluid has been exposed to the reaction zone for a predetermined amount of time.
Claim 13
Independent Claim
189 words
A method for determining in a sample fluid the presence of an analyte which is a member of a specifically binding pair, said method comprising: immersing a port in an otherwise closed receptacle beneath the surface of said sample fluid to entrap a fixed amount of gas within the receptacle, said fluid sample being held in a reservoir which is located inside an enclosure; manipulating the pressure within said enclosure to compress said fixed amount of entrapped gas and to transfer the sample fluid and a labelled reagent which is a member of a specifically binding pair between the reservoir and the receptacle to expose a reaction zone comprising an immobilized substance capable of reacting by non-covalent binding with said analyte to said fluid so that the amount of labelled reagent bound within the reaction zone is proportional to the amount of analyte present; and determining the extent of reaction between the analyte and the substance bound within the reaction zone by observing the amount of label bound within the reaction zone after the sample fluid has been exposed to the reaction zone for a predetermined amount of time.
Claim 14
Dependent Claim
26 words
An assay as in claims 9 or 10, wherein the sample fluid and the labelled reagent are mixed prior to being introduced simultaneously into the receptacle.
Claim 15
Dependent Claim
22 words
An assay as in claims 9 or 10, wherein the sample fluid and the labelled reagent are introduced separately into the receptacle.
Claim 16
Dependent Claim
42 words
An assay as in claims 9 or 10, wherein the analyte and the labelled reagent comprise the same member of the specifically binding pair and compete for binding to the reaction zone which comprises the other member of the specifically binding pair.
Claim 17
Dependent Claim
44 words
An assay as in claims 9 or 10, wherein the labelled reagent and the reaction zone comprise the same member of the specifically binding pair while the analyte comprises the complementary member, and wherein the analyte is introduced prior to introducing the labelling reagent.
Claim 18
Dependent Claim
156 words
In a method for determining an analyte in a sample fluid, wherein said sample fluid is introduced into a solid phase reaction zone and removed from said zone before measuring a signal related to the concentration of analyte, an improved method for transferring the fluid to and from said reaction zone, said method comprising: immersing a port of a single port receptacle beneath the surface of a pool of the sample fluid held in a reservoir located in an enclosure so that a fixed amount of gas is entrapped in the receptacle, where at least one of said receptacle and said reservoir includes the reaction zone in its interior; and manipulating the pressure in the enclosure to compress said fixed amount of entrapped gas and to transfer the sample fluid between the reservoir and the receptacle so that a predetermined amount of sample fluid can be introduced to and withdrawn from the solid phase reaction zone.
Claim 19
Dependent Claim
156 words
In a method for determining an analyte in a sample fluid, wherein said sample fluid is introduced into a solid phase reaction zone and removed from said zone before measuring a signal related to the concentration of analyte, an improved method for transferring the fluid to and from said reaction zone, said method comprising: immersing a port of a single port receptacle beneath the surface of a pool of the sample fluid held in a reservoir located in an enclosure so that a fixed amount of gas is entrapped in the receptacle, where at least one of said receptacle and said reservoir includes the reaction zone in its interior; and manipulating the pressure in the enclosure to compress said fixed amount of entrapped gas and to transfer the sample fluid between the reservoir and the receptacle so that a predetermined amount of sample fluid can be introduced to and withdrawn from the solid phase reaction zone.
Claim 20
Dependent Claim
62 words
An improved method for determining an analyte in a fluid as in claim 15, wherein the reaction zone is present in the receptacle and the pressure is first elevated a predetermined amount to cause a desired volume of the fluid to enter said reaction zone and thereafter lowered to substantially drain the reaction zone prior to determining the extent of reaction therein.
Claim 21
Dependent Claim
42 words
An improved method for determining an analyte as in claim 15, wherein the reaction zone is present in the reservoir and the pressure is elevated to remove unbound label from said reaction zone prior to determining the extent of the reaction therein.
Claim 22
Dependent Claim
198 words
An assay for simultaneously determining an analyte in a plurality of discrete sample fluid reservoirs, said analyte being a member of a specifically bindable pair, said assay comprising: (a) immersing a port in a receptacle beneath the respective surface of each of the sample fluid reservoirs so that a fixed amount of gas is entrapped in said receptacle, each receptacle having a single port and substantially identical dimensions and including a reaction zone characterized by a reaction system which includes a member of the specifically binding pair bound within the reaction zone and produces a detectable signal related to the presence of analyte and a labelled reagent comprising a member of the specifically binding pair in the sample; (b) simultaneously elevating the pressure on the surface of each of the samples to compress said fixed amount of entrapped gas so that a substantially identical amount of fluid enters each receptacle and contacts the reaction zone to complete the reaction system; (c) lowering the pressure so that the fluid is discharged from each reaction zone and the reaction systems produce detectable signals related to the amount of analyte initially present in each sample fluid; and (d) measuring said signals.
Claim 23
Dependent Claim
198 words
An assay for simultaneously determining an analyte in a plurality of discrete sample fluid reservoirs, said analyte being a member of a specifically bindable pair, said assay comprising: (a) immersing a port in a receptacle beneath the respective surface of each of the sample fluid reservoirs so that a fixed amount of gas is entrapped in said receptacle, each receptacle having a single port and substantially identical dimensions and including a reaction zone characterized by a reaction system which includes a member of the specifically binding pair bound within the reaction zone and produces a detectable signal related to the presence of analyte and a labelled reagent comprising a member of the specifically binding pair in the sample; (b) simultaneously elevating the pressure on the surface of each of the samples to compress said fixed amount of entrapped gas so that a substantially identical amount of fluid enters each receptacle and contacts the reaction zone to complete the reaction system; (c) lowering the pressure so that the fluid is discharged from each reaction zone and the reaction systems produce detectable signals related to the amount of analyte initially present in each sample fluid; and (d) measuring said signals.
Claim 24
Dependent Claim
22 words
An assay as in claim 18, further comprising repeating steps (a) through (d) in order to periodically determine the extent of reaction.
Claim 25
Dependent Claim
35 words
An assay as in claim 18, wherein the pressure on the surface of each of the samples is maintained uniform by keeping the samples in a common plenum and changing the pressure in the plenum.
Claim 26
Dependent Claim
24 words
An assay as in claim 20, wherein the pressure in the plenum is periodically varied to agitate the sample fluid in the reaction zone.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1987-10-16 | 1988-04-16 | 1988-10-16 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-10-16 | 1992-04-16 | 1992-10-16 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-10-16 | 1996-04-16 | 1996-10-16 | $7700.00 | Window Closed / Lapsed |