title: An Assay of Measuring Platelet Reactivity Using Monoclonal Antibody against Activated Platelet Glycoprotein IIb/IIIa in Patients Taking Clopidogrel.
abstract: OBJECTIVE
  Residual platelet reactivity in patients who are taking clopidogrel is commonly measured with VerifyNow assay, which is based on the principle of light transmission aggregometry. However, to evaluate the residual platelet reactivity, it would be more accurate if the reactivity of platelet glycoprotein (GP) IIb/IIIa is directly monitored. In this study, PAC1, a monoclonal antibody against activated platelet GP IIb/IIIa, was used to measure the residual platelet reactivity.
  
  
  METHODS
  Twenty seven patients with coronary artery disease taking clopidogrel were enrolled. Platelets in whole blood were stained with fluorescein isothiocyanate (FITC)-conjugated PAC1. Mean fluorescence intensity (MFI) and % positive platelets (PP) were measured with flow cytometry, and the binding index (BI; MFI × %PP/100) was calculated. P2Y12 reaction unit (PRU) and % inhibition of VerifyNow assay were also measured in the usual manner.
  
  
  RESULTS
  PRU of VerifyNow assay correlated significantly with MFI, %PP, and BI at 10 µM (r=0.59, 0.73, and 0.60, respectively, all p<0.005) and 20 µM of adenosine diphosphate (ADP; r=0.61, 0.75, and 0.63, respectively, all p<0.005). The % inhibition also correlated significantly with MFI, %PP, and BI at 10 µM (r=-0.60, -0.69, and -0.59, respectively, all p<0.005) and 20 µM of ADP (r=-0.63, -0.71, and -0.62, respectively, all p<0.005).
  
  
  CONCLUSIONS
  Direct measurements of the reactivity of platelet GP IIb/IIIa were feasible using PAC1 and flow cytometry in patients taking clopidogrel. Further clinical studies are required to determine the cut-off values which would define high residual platelet reactivity in patients on this treatment protocol.
authors.affiliation: Department of Cardiology, Jeju National University Hospital, Jeju, Korea.
authors.forename: Seung-Jae
authors.initials: SJ
authors.lastname: Joo
authors.affiliation: Department of Cardiology, Jeju National University Hospital, Jeju, Korea.
authors.forename: Joon-Hyouk
authors.initials: JH
authors.lastname: Choi
authors.affiliation: Department of Cardiology, Jeju National University Hospital, Jeju, Korea.
authors.forename: Song-Yi
authors.initials: SY
authors.lastname: Kim
authors.affiliation: Department of Cardiology, Jeju National University Hospital, Jeju, Korea.
authors.forename: Ki-Seok
authors.initials: KS
authors.lastname: Kim
authors.affiliation: Department of Laboratory Medicine, Jeju National University Hospital, Jeju, Korea.
authors.forename: Young Ree
authors.initials: YR
authors.lastname: Kim
authors.affiliation: Department of Laboratory Medicine, Jeju National University Hospital, Jeju, Korea.
authors.forename: Sung Ha
authors.initials: SH
authors.lastname: Kang
keywords: Blood platelets; Clopidogrel; Flow cytometry; Glycoprotein IIb/IIIa; Platelet function test