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US 2025/0076264 A1Published application
Point of care testing system for antithrombin iii
AssigneeUniversity of Florida Research Foundation, Inc.
InventorsAdam S. Veige, Yong Zeng, Bruce Davis Spiess, Amanda Orta Santana, He Yan
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Abstract
Compounds and compositions for determining the level of antithrombin (ATIII) in a sample are described along with a system for determining the level of ATIII in a point-of-care setting. Methods of forming the compounds and compositions are also described. Methods of using the compounds and compositions to quantify the level of ATIII in a subject are further described. A system and apparatus are provided that determine the level of ATIII in a point-of-care setting in an efficient manner to facilitate determining a dosage or heparin or ATIII to administer to a patient.
Claims27
A system for determining a level of anti-thrombin (ATIII) in a subject comprising:
- a column defining an inlet and an outlet, wherein heparin beads are retained within the column;
- a manifold connected to the inlet of the column, wherein the manifold comprises a plurality of ports, each port configured to receive a fluid, wherein the manifold conducts the fluid from each port to the inlet of the column;
- a first syringe comprising a first washing buffer received at a first port of the plurality of ports of the manifold;
- a second syringe comprising plasma received at a second port of the plurality of ports of the manifold;
- a third syringe comprising a second washing buffer received at a third port of the plurality of ports of the manifold;
- and
- a fourth syringe comprising an aptamer-thrombin complex received at a fourth port of the plurality of ports of the manifold, wherein the aptamer-thrombin complex comprises a first signaling aptamer comprising a first thrombin-specific aptamer, a first hybridization sequence, and a fluorescent label, a second signaling aptamer comprising a second thrombin-specific aptamer, a second hybridization sequence, and a quencher and thrombin, wherein the first and second hybridization sequences are complementary to each other, and wherein fluorescence of the fluorescent label is quenched when the first and second thrombin-specific signaling aptamers are bound to the thrombin.
4 more elements
A method for determining a level of anti-thrombin (ATIII) in a subject comprising:
- driving a first washing buffer through a column within which are suspended heparin beads;
- driving a sample through the column, wherein the sample comprises a plasma from the subject;
- driving a second washing buffer through the column;
- driving a solution comprising an aptamer-thrombin complex through the column, wherein the aptamer-thrombin complex comprises a first signaling aptamer comprising a first thrombin-specific aptamer, a first hybridization sequence, and a fluorescent label, a second signaling aptamer comprising a second thrombin-specific aptamer, a second hybridization sequence, and a quencher and thrombin, wherein the first and second hybridization sequences are complementary to each other, and wherein fluorescence of the fluorescent label is quenched when the first and second thrombin-specific signaling aptamers are bound to the thrombin;
- and
- collecting an eluent from the column, wherein the eluent comprises the aptamer-thrombin complex and ATIII from the sample
- measuring fluorescence in the solution, wherein the level of fluorescence is indicative of a level of ATIII in the plasma.
4 more elements
An apparatus for determining a level of anti-thrombin (ATIII) in a subject comprising:
- a column defining an inlet and an outlet;
- a manifold defining a plurality of ports and a manifold outlet, wherein the manifold outlet is connected to the inlet of the column;
- a substrate defining a waste flow path extending between a substrate inlet and a waste reservoir, and a fluorescence flow path extending between the substrate inlet and a fluorescence reservoir;
- a waste valve configured to open and close the waste flow path between the substrate inlet and the waste reservoir;
- and
- a fluorescence valve configured to open and close the fluorescence flow path between the substrate inlet and the fluorescence reservoir.
3 more elements
A kit for determining the level of anti-thrombin (ATIII) in a subject, comprising:
- (a) a first signaling aptamer comprising a first thrombin-specific aptamer, a first hybridization sequence, and a fluorescent label;
- (b) a second signaling aptamer comprising a second thrombin-specific aptamer, a second hybridization sequence, and a quencher
- (c) thrombin;
- and
- (d) heparin beads,
- wherein the first and second hybridization sequences are complementary to each other, and wherein fluorescence of the fluorescent label is quenched when the first and second thrombin-specific signaling aptamers are bound to thrombin.
3 more elements
A method of determining a level of ATIII in a subject, comprising:
- (a) contacting a sample from the subject with heparin linked to a support, wherein any ATIII in the sample binds to the heparin;
- (b) purifying the sample to remove one or more components of the sample that do not bind to the heparin to form a purified sample;
- (c) contacting the purified sample with an aptamer-thrombin complex to form an assay solution, wherein the aptamer-thrombin complex comprises a first signaling aptamer comprising a first thrombin-specific aptamer, a first hybridization sequence, and a fluorescent label, a second signaling aptamer comprising a second thrombin-specific aptamer, a second hybridization sequence, and a quencher and thrombin, wherein the first and second hybridization sequences are complementary to each other, and wherein fluorescence of the fluorescent label is quenched when the first and second thrombin-specific signaling aptamers are bound to the thrombin;
- and
- (d) measuring fluorescence in the assay solution wherein the level of fluorescence is proportional to the level of ATIII in the sample.
2 more elements
Dependencies are inferred from the claim text.