PMID- 20508902
OWN - NLM
STAT- MEDLINE
DCOM- 20101202
LR  - 20151119
IS  - 2567-689X (Electronic)
IS  - 0340-6245 (Linking)
VI  - 104
IP  - 2
DP  - 2010 Aug
TI  - Development of a high-throughput ELISA assay for platelet function testing using 
      platelet-rich plasma or whole blood.
PG  - 392-401
LID - 10.1160/TH09-07-0505 [doi]
AB  - Platelets play an essential role in the development of cardiovascular diseases 
      and are the target of several agents that can inhibit their function. Despite the 
      existence of a wide array of techniques to study platelet function, an assay to 
      evaluate several platelet signalling pathways in a high-throughput fashion, 
      combined with minimal blood volume and handling is still needed. We have 
      developed a sensitive assay in the form of a sandwich ELISA where monoclonal 
      antibodies against P-selectin or alphaIIbbeta3 and GPIbalpha were used to capture 
      and detect platelets, respectively, in the presence of five different agonists 
      [ADP, TRAP (thrombin receptor agonist), U46619 (thromboxane A2 analogue), 
      collagen-related-peptide, and arachidonic acid]. Binding of platelets to the 
      antibodies increased dose-dependently with the concentration of either agonist, 
      while binding of ADP-activated platelets was abrogated when inhibitors of 
      platelet activation were concomitantly added. The test showed good sample 
      reproducibility in 15 healthy donors with conserved platelet response to agonists 
      throughout the assay. Healthy subjects could be identified as normal-, hypo- or 
      hyper-responders for each agonist, which for most cases (73%) was confirmed upon 
      retesting. Finally, we demonstrated that the platelet ELISA assay can not only be 
      used in platelet-rich plasma but also in whole blood; it now awaits large scale 
      studies to assess its full screening and diagnostic values.
FAU - Salles, Isabelle
AU  - Salles I
AD  - Laboratory for Thrombosis Research, KU Leuven-Campus Kortrijk, Kortrijk, Belgium. 
      i.salles@imperial.ac.uk
FAU - Broos, Katleen
AU  - Broos K
FAU - Fontayne, Alexandre
AU  - Fontayne A
FAU - Szanto, Timea
AU  - Szanto T
FAU - Ruan, Changgeng
AU  - Ruan C
FAU - Nurden, Alan T
AU  - Nurden AT
FAU - Vanhoorelbeke, Karen
AU  - Vanhoorelbeke K
FAU - Deckmyn, Hans
AU  - Deckmyn H
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20100527
PL  - Germany
TA  - Thromb Haemost
JT  - Thrombosis and haemostasis
JID - 7608063
RN  - 0 (A1BG protein, human)
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Carrier Proteins)
RN  - 0 (Glycoproteins)
RN  - 0 (ITGA2B protein, human)
RN  - 0 (ITGB3 protein, human)
RN  - 0 (Immunoglobulins)
RN  - 0 (Integrin alpha2)
RN  - 0 (Integrin beta3)
RN  - 0 (P-Selectin)
RN  - 0 (Peptides)
RN  - 0 (Platelet Aggregation Inhibitors)
RN  - 0 (collagen-related peptide)
RN  - 27YG812J1I (Arachidonic Acid)
RN  - 61D2G4IYVH (Adenosine Diphosphate)
RN  - 76898-47-0 (15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid)
RN  - 79032-48-7 (S-Nitroso-N-Acetylpenicillamine)
RN  - F5TD010360 (Alprostadil)
SB  - IM
MH  - 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid
MH  - Adenosine Diphosphate
MH  - Alprostadil/pharmacology
MH  - Antibodies, Monoclonal
MH  - Arachidonic Acid
MH  - Blood Platelets/drug effects/immunology/physiology
MH  - Carrier Proteins
MH  - Dose-Response Relationship, Drug
MH  - *Enzyme-Linked Immunosorbent Assay
MH  - Flow Cytometry
MH  - Glycoproteins/immunology
MH  - *High-Throughput Screening Assays
MH  - Humans
MH  - Immunoglobulins/immunology
MH  - Integrin alpha2/immunology
MH  - Integrin beta3/immunology
MH  - Observer Variation
MH  - P-Selectin/immunology
MH  - Peptides
MH  - *Platelet Activation/drug effects
MH  - Platelet Aggregation Inhibitors/pharmacology
MH  - *Platelet Function Tests
MH  - Predictive Value of Tests
MH  - Reproducibility of Results
MH  - S-Nitroso-N-Acetylpenicillamine/pharmacology
EDAT- 2010/05/29 06:00
MHDA- 2010/12/14 06:00
CRDT- 2010/05/29 06:00
PHST- 2009/07/31 00:00 [received]
PHST- 2010/03/19 00:00 [accepted]
PHST- 2010/05/29 06:00 [entrez]
PHST- 2010/05/29 06:00 [pubmed]
PHST- 2010/12/14 06:00 [medline]
AID - 09-07-0505 [pii]
AID - 10.1160/TH09-07-0505 [doi]
PST - ppublish
SO  - Thromb Haemost. 2010 Aug;104(2):392-401. doi: 10.1160/TH09-07-0505. Epub 2010 May 
      27.