Reversible flow chromatographic binding assay
First Claim
1. An assay system for performing an assay which determines the presence or quantity of an analyte in a fluid sample suspected of containing the analyte by detecting analyte bound to an immobilized analyte capture reagent after unbound material has been washed away, the system comprising:
- (a) an elongated solid phase flow matrix capable of driving fluid movement, the flow matrix comprising,(i) a first matrix region for receipt of an analyte-containing fluid,(ii) a second matrix region at which the analyte capture reagent is immobilized, the second matrix region being positioned in between the first matrix region and a third matrix region (iii),(iii) the third matrix region being for receipt of liquid capable of removing from the second matrix region substances that are not bound to the analyte capture reagent; and
(iv) an absorbent reservoir positioned to contact the matrix at an absorption region, the second matrix region being between the absorption region and the third matrix region;
(b) a mobile analyte binding reagent which is capable of;
i) binding to analyte;
ii) moving through the matrix; and
iii) facilitating detection of analyte bound to the second matrix region, said mobile analyte binding reagent comprising an analyte binding component and a detectable label;
whereby operation of the system initially causes movement of analyte-containing fluid in a first direction away from the first matrix region through the second matrix region; and
, after introduction of the liquid into the third region of the flow matrix and the absorbent reservoir is in contact with the absorption region of the matrix, flow in the matrix occurs in a second direction opposite the first direction, the second direction being away from the third matrix region and into the second region, whereby the analyte containing fluid reverses flow and does not flow in said first direction past the third matrix region.
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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.
103 Citations
3 Claims
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1. An assay system for performing an assay which determines the presence or quantity of an analyte in a fluid sample suspected of containing the analyte by detecting analyte bound to an immobilized analyte capture reagent after unbound material has been washed away, the system comprising:
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(a) an elongated solid phase flow matrix capable of driving fluid movement, the flow matrix comprising, (i) a first matrix region for receipt of an analyte-containing fluid, (ii) a second matrix region at which the analyte capture reagent is immobilized, the second matrix region being positioned in between the first matrix region and a third matrix region (iii), (iii) the third matrix region being for receipt of liquid capable of removing from the second matrix region substances that are not bound to the analyte capture reagent; and (iv) an absorbent reservoir positioned to contact the matrix at an absorption region, the second matrix region being between the absorption region and the third matrix region; (b) a mobile analyte binding reagent which is capable of; i) binding to analyte; ii) moving through the matrix; and iii) facilitating detection of analyte bound to the second matrix region, said mobile analyte binding reagent comprising an analyte binding component and a detectable label; whereby operation of the system initially causes movement of analyte-containing fluid in a first direction away from the first matrix region through the second matrix region; and
, after introduction of the liquid into the third region of the flow matrix and the absorbent reservoir is in contact with the absorption region of the matrix, flow in the matrix occurs in a second direction opposite the first direction, the second direction being away from the third matrix region and into the second region, whereby the analyte containing fluid reverses flow and does not flow in said first direction past the third matrix region. - View Dependent Claims (2, 3)
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Specification