PMID- 20823666
OWN - NLM
STAT- MEDLINE
DCOM- 20110118
LR  - 20191210
IS  - 1349-3329 (Electronic)
IS  - 0040-8727 (Linking)
VI  - 222
IP  - 1
DP  - 2010 Sep
TI  - A high-throughput biotin-avidin-ELISA for studying expression of platelet 
      membrane glycoproteins and its clinical application.
PG  - 83-8
AB  - Platelet membrane glycoproteins (GPs) are critical for normal platelet adhesion, 
      activation and aggregation. To define the abnormalities in surface GP expression 
      on circulating platelets and provide a better biomarker of bleeding and 
      thrombotic disorders, we have developed a accurate, time-saving and 
      high-throughput biotin-avidin enzyme-linked immunosorbent assay (BA-ELISA) with 
      the monoclonal antibodies (mAbs), 7E3 against the complex of GPIIb and GPIIIa 
      (GPIIb/IIIa), SZ-51 against P-selectin, and SZ-2 against GPIb, respectively. The 
      levels of P-selectin and GPIIb/IIIa were measured in patients with acute 
      myocardial infarction (AMI), intracerebral hemorrhage (ICH), or diabetes mellitus 
      (DM)) and healthy subjects. Inhibition of GP expression was evaluated with SZ-21, 
      an inhibitory mAb against GPIIIa and aspirin, respectively. The sensitivity of 
      BA-ELISA is high enough to detect platelet count as low as 3.13 x 10(9)/L in 
      platelet-rich plasma (PRP). Both the inter-assay and intra-assay coefficient 
      variation are less than 10%. Adenosine diphosphate (ADP)-induced or 
      non-ADP-induced expression of P-selectin and GPIIb/IIIa was significantly higher 
      in AMI, ICH or DM than that in controls (P < 0.01 for each). Either SZ-21 or 
      aspirin can inhibit the ADP-induced expression of P-selectin and GPIIb/IIIa. 
      Importantly, a high correlation was detected between BA-ELISA and flow cytometry 
      methods. These observations indicate that BA-ELISA is a sensitive and 
      high-throughput assay for evaluating platelet GP expression. The newly developed 
      BA-ELISA can be popularized in community hospitals, because it does not require 
      sophisticated equipments and reagents. This method is suitable for screening 
      inhibitors of platelet activation and has a potential in use for diagnostic 
      purposes.
FAU - Zhang, Youtao
AU  - Zhang Y
AD  - Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow 
      University, Key Laboratory of Thrombosis and Haemostasis, Ministry of Health, 
      Suzhou, Jiangsu, China.
FAU - Zhao, Yiming
AU  - Zhao Y
FAU - Lu, Shiqi
AU  - Lu S
FAU - Zhu, Mingqing
AU  - Zhu M
FAU - He, Yang
AU  - He Y
FAU - Ruan, Changgeng
AU  - Ruan C
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Japan
TA  - Tohoku J Exp Med
JT  - The Tohoku journal of experimental medicine
JID - 0417355
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Biomarkers)
RN  - 0 (Platelet Membrane Glycoproteins)
RN  - 1405-69-2 (Avidin)
RN  - 61D2G4IYVH (Adenosine Diphosphate)
RN  - 6SO6U10H04 (Biotin)
SB  - IM
MH  - Adenosine Diphosphate
MH  - Antibodies, Monoclonal
MH  - Avidin
MH  - Biomarkers/*blood
MH  - Biotin
MH  - Cerebral Hemorrhage/diagnosis/metabolism
MH  - Diabetes Mellitus/diagnosis/metabolism
MH  - Enzyme-Linked Immunosorbent Assay/*methods
MH  - Flow Cytometry
MH  - Humans
MH  - Myocardial Infarction/diagnosis/metabolism
MH  - Platelet Membrane Glycoproteins/*analysis/*metabolism
MH  - Sensitivity and Specificity
EDAT- 2010/09/09 06:00
MHDA- 2011/01/19 06:00
CRDT- 2010/09/09 06:00
PHST- 2010/09/09 06:00 [entrez]
PHST- 2010/09/09 06:00 [pubmed]
PHST- 2011/01/19 06:00 [medline]
AID - JST.JSTAGE/tjem/222.83 [pii]
AID - 10.1620/tjem.222.83 [doi]
PST - ppublish
SO  - Tohoku J Exp Med. 2010 Sep;222(1):83-8. doi: 10.1620/tjem.222.83.