Reversible flow chromatographic binding assay
First Claim
1. A device for performing an assay which determines the presence or quantity of an analyte in a fluid sample by detecting binding of said analyte to at least one immobilized analyte capture reagent after washing unbound material from said immobilized analyte capture reagent, said device comprising:
- (a) an elongated solid phase flow matrix, said flow matrix comprising capillary channels capable of driving capillary fluid movement, said flow matrix further comprisingi) a first region adapted for receipt of said fluid sample,ii) a second region at which said analyte capture reagent is immobilized,iii) a third region for application of a liquid detector reagent capable of removing unbound substances from said second region;
said second region being positioned intermediate to said first region and said third region;
(b) an absorbent reservoir of high volume capacity, wherein, prior to use of said device, said absorbent reservoir is not in fluidic contact with said flow matrix;
said device further comprising means for establishing fluidic contact between said absorbent reservoir and said flow matrix at a position selected so that said second region is between said absorbent reservoir and said third region;
(c) a sealed container of said liquid detector reagent positioned to be introduced at said third region of said matrix;
whereby said flow matrix and said regions thereof are sized and positioned to cause said fluid sample to flow initially along said elongated flow matrix in one direction toward and through said second region, and subsequently, upon introduction of said liquid detector reagent into the third region of said flow matrix, said liquid detector reagent to flow along said elongated flow matrix in a second direction opposite said first direction, through said second region, and into said absorbent reservoir, drawing unbound substances with it.
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Accused Products
Abstract
Disclosed are devices and methods for performing an assay which determines the presence or quantity of an analyte in a fluid sample by detecting binding of the analyte to at least one immobilized analyte capture reagent and washing unbound material from the immobilized analyte capture reagent. The devices and methods an elongated solid phase flow matrix, including capillary channels capable of driving capillary fluid movement, and further including (i) a first region adapted for receipt of the fluid sample, (ii) a second region at which the analyte capture reagent is immobilized, (iii) a third region for application of a wash reagent capable of removing unbound substances from the second region; and (iv) an absorbent reservoir of high volume capacity. The second region is positioned intermediate to the first region and the third region and intermediate to the absorbent reservoir and the third region. The device also includes means to detect analyte bound at the second region. The flow matrix and the regions thereof are sized and positioned to cause the fluid sample to flow initially along the elongated flow matrix in one direction toward and through the second region, and subsequently, the liquid reagent to flow along the elongated flow matrix in a second direction opposite the first direction, through the second region, and into the absorbent reservoir, drawing unbound substances with it.
320 Citations
36 Claims
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1. A device for performing an assay which determines the presence or quantity of an analyte in a fluid sample by detecting binding of said analyte to at least one immobilized analyte capture reagent after washing unbound material from said immobilized analyte capture reagent, said device comprising:
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(a) an elongated solid phase flow matrix, said flow matrix comprising capillary channels capable of driving capillary fluid movement, said flow matrix further comprising i) a first region adapted for receipt of said fluid sample, ii) a second region at which said analyte capture reagent is immobilized, iii) a third region for application of a liquid detector reagent capable of removing unbound substances from said second region; said second region being positioned intermediate to said first region and said third region; (b) an absorbent reservoir of high volume capacity, wherein, prior to use of said device, said absorbent reservoir is not in fluidic contact with said flow matrix;
said device further comprising means for establishing fluidic contact between said absorbent reservoir and said flow matrix at a position selected so that said second region is between said absorbent reservoir and said third region;(c) a sealed container of said liquid detector reagent positioned to be introduced at said third region of said matrix; whereby said flow matrix and said regions thereof are sized and positioned to cause said fluid sample to flow initially along said elongated flow matrix in one direction toward and through said second region, and subsequently, upon introduction of said liquid detector reagent into the third region of said flow matrix, said liquid detector reagent to flow along said elongated flow matrix in a second direction opposite said first direction, through said second region, and into said absorbent reservoir, drawing unbound substances with it. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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19. A method for performing an assay which determines the presence or quantity of an analyte in a fluid sample by detecting binding of said analyte to at least one immobilized analyte capture reagent after washing unbound material from said immobilized analyte capture reagent, said method comprising:
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(a) providing (i) an elongated solid phase flow matrix, said flow matrix comprising capillary channels capable of driving capillary fluid movement, said flow matrix further comprising a) a first region adapted for receipt of said fluid sample, b) a second region at which said analyte capture reagent is immobilized, c) a third region for application of a liquid detector reagent capable of removing unbound substances from said second region;
said second region being positioned intermediate to said first region and said third region;(ii) an absorbent reservoir of high volume capacity, wherein, prior to performing said method, said absorbent reservoir is not in fluidic contact with said flow matrix; (b) applying said fluid sample to said first region of said flow matrix; (c) allowing said fluid sample to flow in a first direction through said second region, and then introducing said liquid detector reagent into said flow matrix at said third region; (d) moving said absorbent reservoir into fluidic contact with said flow matrix, such that said sample and said liquid detector reagent flow in a second direction, opposite to said first direction; and (e) detecting said analyte bound at said second region. - View Dependent Claims (20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33)
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34. A device for performing an assay which determines the presence or quantity of an analyte in a fluid sample by detecting binding of said analyte to at least one immobilized analyte capture reagent after washing unbound material from said immobilized analyte capture reagent, said device comprising:
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(a) an elongated fluid flow matrix comprising, (1) a first segment for receiving a fluid sample, (2) a second region at which analyte capture reagents are immobilized, (3) a third region for application of a liquid detector reagent capable of removing unbound substances from said second region, (b) an absorbent reservoir of high volume capacity, (c) a sealed container of said liquid detector reagent positioned to be introduced at said third region of said flow matrix, (d) at least one soluble barrier positioned to block flow of said liquid detector reagent from said container to said absorbent reservoir, said second region being positioned intermediate to said third region and said absorbent reservoir, said soluble barrier blocking flow of said liquid detector reagent to said absorbent reservoir until after said sample has flowed from said first region through said second region, at which point said barrier dissolves permitting said liquid detector reagent to flow in said wash path. - View Dependent Claims (35, 36)
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Specification