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Monitoring modulators of platelet aggregation in a microtiter plate assay

Monitoring modulators of platelet aggregation in a microtiter plate assay
Monitoring modulators of platelet aggregation in a microtiter plate assay
Platelets play a central role in maintaining biological hemostasis. Inappropriate platelet activation is responsible for thrombotic diseases such as myocardial infarction and stroke. Therefore, novel agents that can inhibit platelet activation are necessary. However, assays that monitor platelet aggregation are generally time-consuming and require high volumes of blood and specialized equipment. Therefore, a medium- to high-throughput assay that can monitor platelet aggregation would be considered useful. Such an assay should be sensitive, comparable to the "gold standard" assay of platelet aggregometry, and able to monitor multiple samples simultaneously but with low assay volumes. We have developed such a microtiter assay. It can assay an average of 60 independent treatments per 60 ml blood donation and demonstrates greater sensitivity than the current gold standard assay, namely platelet aggregation in stirring conditions in a platelet aggregometer. The microtiter plate (MTP) assay can detect known inhibitors of platelet function such as indomethacin, aspirin, and ReoPro. It is highly reproducible when using standard doses of agonists such as thrombin receptor-activating peptide (20 microM) and collagen (0.19 mg/ml). Finally, the MTP assay is rapid and sensitive and can detect unknown platelet-modulating agents from a library of compounds.

platelet, aggregation, peptide, inhibitors, microtiter assay
0003-2697
77-84
Moran, N.
4ea87433-6098-4b97-8098-fe43c722d9c4
Kiernan, A.
9b736a0f-dff9-497d-82ac-e50a06de8397
Dunnan, E.
d19d497e-fbba-4e0c-8ebc-6b62a1a4d350
Edwards, Richard J.
9d25e74f-dc0d-455a-832c-5f363d864c43
Shields, D.C.
57ffee4f-0277-4b3d-9c7a-8c328637d8e6
Kenny, D.
78c0def1-b345-4ddf-8a76-4b5537bf350a
Moran, N.
4ea87433-6098-4b97-8098-fe43c722d9c4
Kiernan, A.
9b736a0f-dff9-497d-82ac-e50a06de8397
Dunnan, E.
d19d497e-fbba-4e0c-8ebc-6b62a1a4d350
Edwards, Richard J.
9d25e74f-dc0d-455a-832c-5f363d864c43
Shields, D.C.
57ffee4f-0277-4b3d-9c7a-8c328637d8e6
Kenny, D.
78c0def1-b345-4ddf-8a76-4b5537bf350a

Moran, N., Kiernan, A., Dunnan, E., Edwards, Richard J., Shields, D.C. and Kenny, D. (2006) Monitoring modulators of platelet aggregation in a microtiter plate assay. Analytical Biochemistry, 357 (1), 77-84. (doi:10.1016/j.ab.2006.06.037).

Record type: Article

Abstract

Platelets play a central role in maintaining biological hemostasis. Inappropriate platelet activation is responsible for thrombotic diseases such as myocardial infarction and stroke. Therefore, novel agents that can inhibit platelet activation are necessary. However, assays that monitor platelet aggregation are generally time-consuming and require high volumes of blood and specialized equipment. Therefore, a medium- to high-throughput assay that can monitor platelet aggregation would be considered useful. Such an assay should be sensitive, comparable to the "gold standard" assay of platelet aggregometry, and able to monitor multiple samples simultaneously but with low assay volumes. We have developed such a microtiter assay. It can assay an average of 60 independent treatments per 60 ml blood donation and demonstrates greater sensitivity than the current gold standard assay, namely platelet aggregation in stirring conditions in a platelet aggregometer. The microtiter plate (MTP) assay can detect known inhibitors of platelet function such as indomethacin, aspirin, and ReoPro. It is highly reproducible when using standard doses of agonists such as thrombin receptor-activating peptide (20 microM) and collagen (0.19 mg/ml). Finally, the MTP assay is rapid and sensitive and can detect unknown platelet-modulating agents from a library of compounds.

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More information

Published date: 14 July 2006
Keywords: platelet, aggregation, peptide, inhibitors, microtiter assay

Identifiers

Local EPrints ID: 143467
URI: http://eprints.soton.ac.uk/id/eprint/143467
ISSN: 0003-2697
PURE UUID: 9bb2ff7a-2c7b-4574-9a1a-73c0fe4c7d05

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Date deposited: 21 Jun 2010 10:58
Last modified: 14 Mar 2024 00:43

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Contributors

Author: N. Moran
Author: A. Kiernan
Author: E. Dunnan
Author: Richard J. Edwards
Author: D.C. Shields
Author: D. Kenny

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