Identifying human platelet glycoproteins IIb and IIla by capillary electrophoresis

G. Vecchione, M. Margaglione, E. Grandone, D. Colaizzo, G. Cappucci, N. Giuliani, M. d'Addedda, G. D'Andrea, M. Nobile, A. Amoriello, G. Di Minno

Research output: Contribution to journalArticle

Abstract

Glanzmann thrombasthenia (GT) is an inherited hemorrhagic defect due to a failure of the platelet membrane glycoprotein (GP) IIb-IIIa complex. Capillary electrophoresis (CE) analysis of solubilized platelet membranes from normal individuals showed the presence of two peaks with a migration time of 27 and 29 min, respectively. An excellent run-to-run and day-to-day reproducibility of the technique (<1% variation of the retention time) was documented. Using an automated Ferguson method, the apparent molecular masses were 100.0 kDa and 138.5 kDa, respectively. Immunoprecipitation with monoclonal antibodies anti-GP IIIa (B59.2.1) and anti-IIb (61.9.1.3) showed the two peaks as IIIa and IIb, respectively. Electropherograms of a GT young man showed the lack of both peaks. Less than 50% of each peak was present in his parents. Polyacrylamide gel electrophoresis (PAGE), immunoblotting, and flow cytometry analyses showed that GP IIb and IIIa were undetectable in the platelet membranes from the propositus, half of the normal amount being present in both parents. These findings indicate CE to be a rapid, sensitive and reliable tool to investigate patients with abnormalities of the GP IIb-IIIa complex.

Original languageEnglish
Pages (from-to)1468-1474
Number of pages7
JournalElectrophoresis
Volume19
Issue number8-9
DOIs
Publication statusPublished - 1998

Fingerprint

Platelet Membrane Glycoproteins
Capillary electrophoresis
Platelet Glycoprotein GPIIb-IIIa Complex
Capillary Electrophoresis
Thrombasthenia
Platelets
Blood Platelets
Parents
Platelet Membrane Glycoprotein IIb
Integrin beta3
Membranes
Flow cytometry
Molecular mass
Electrophoresis
Immunoprecipitation
Immunoblotting
Polyacrylamide Gel Electrophoresis
Glycoproteins
Flow Cytometry
Monoclonal Antibodies

Keywords

  • Capillary electrophoresis
  • Glanzmann's thrombasthenia
  • Glycoproteins
  • Platelet membrane

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Identifying human platelet glycoproteins IIb and IIla by capillary electrophoresis. / Vecchione, G.; Margaglione, M.; Grandone, E.; Colaizzo, D.; Cappucci, G.; Giuliani, N.; d'Addedda, M.; D'Andrea, G.; Nobile, M.; Amoriello, A.; Di Minno, G.

In: Electrophoresis, Vol. 19, No. 8-9, 1998, p. 1468-1474.

Research output: Contribution to journalArticle

Vecchione, G, Margaglione, M, Grandone, E, Colaizzo, D, Cappucci, G, Giuliani, N, d'Addedda, M, D'Andrea, G, Nobile, M, Amoriello, A & Di Minno, G 1998, 'Identifying human platelet glycoproteins IIb and IIla by capillary electrophoresis', Electrophoresis, vol. 19, no. 8-9, pp. 1468-1474. https://doi.org/10.1002/elps.1150190842
Vecchione, G. ; Margaglione, M. ; Grandone, E. ; Colaizzo, D. ; Cappucci, G. ; Giuliani, N. ; d'Addedda, M. ; D'Andrea, G. ; Nobile, M. ; Amoriello, A. ; Di Minno, G. / Identifying human platelet glycoproteins IIb and IIla by capillary electrophoresis. In: Electrophoresis. 1998 ; Vol. 19, No. 8-9. pp. 1468-1474.
@article{d4daa4d662834a79bc91ce2d29e59804,
title = "Identifying human platelet glycoproteins IIb and IIla by capillary electrophoresis",
abstract = "Glanzmann thrombasthenia (GT) is an inherited hemorrhagic defect due to a failure of the platelet membrane glycoprotein (GP) IIb-IIIa complex. Capillary electrophoresis (CE) analysis of solubilized platelet membranes from normal individuals showed the presence of two peaks with a migration time of 27 and 29 min, respectively. An excellent run-to-run and day-to-day reproducibility of the technique (<1{\%} variation of the retention time) was documented. Using an automated Ferguson method, the apparent molecular masses were 100.0 kDa and 138.5 kDa, respectively. Immunoprecipitation with monoclonal antibodies anti-GP IIIa (B59.2.1) and anti-IIb (61.9.1.3) showed the two peaks as IIIa and IIb, respectively. Electropherograms of a GT young man showed the lack of both peaks. Less than 50{\%} of each peak was present in his parents. Polyacrylamide gel electrophoresis (PAGE), immunoblotting, and flow cytometry analyses showed that GP IIb and IIIa were undetectable in the platelet membranes from the propositus, half of the normal amount being present in both parents. These findings indicate CE to be a rapid, sensitive and reliable tool to investigate patients with abnormalities of the GP IIb-IIIa complex.",
keywords = "Capillary electrophoresis, Glanzmann's thrombasthenia, Glycoproteins, Platelet membrane",
author = "G. Vecchione and M. Margaglione and E. Grandone and D. Colaizzo and G. Cappucci and N. Giuliani and M. d'Addedda and G. D'Andrea and M. Nobile and A. Amoriello and {Di Minno}, G.",
year = "1998",
doi = "10.1002/elps.1150190842",
language = "English",
volume = "19",
pages = "1468--1474",
journal = "Electrophoresis",
issn = "0173-0835",
publisher = "Wiley-VCH Verlag",
number = "8-9",

}

TY - JOUR

T1 - Identifying human platelet glycoproteins IIb and IIla by capillary electrophoresis

AU - Vecchione, G.

AU - Margaglione, M.

AU - Grandone, E.

AU - Colaizzo, D.

AU - Cappucci, G.

AU - Giuliani, N.

AU - d'Addedda, M.

AU - D'Andrea, G.

AU - Nobile, M.

AU - Amoriello, A.

AU - Di Minno, G.

PY - 1998

Y1 - 1998

N2 - Glanzmann thrombasthenia (GT) is an inherited hemorrhagic defect due to a failure of the platelet membrane glycoprotein (GP) IIb-IIIa complex. Capillary electrophoresis (CE) analysis of solubilized platelet membranes from normal individuals showed the presence of two peaks with a migration time of 27 and 29 min, respectively. An excellent run-to-run and day-to-day reproducibility of the technique (<1% variation of the retention time) was documented. Using an automated Ferguson method, the apparent molecular masses were 100.0 kDa and 138.5 kDa, respectively. Immunoprecipitation with monoclonal antibodies anti-GP IIIa (B59.2.1) and anti-IIb (61.9.1.3) showed the two peaks as IIIa and IIb, respectively. Electropherograms of a GT young man showed the lack of both peaks. Less than 50% of each peak was present in his parents. Polyacrylamide gel electrophoresis (PAGE), immunoblotting, and flow cytometry analyses showed that GP IIb and IIIa were undetectable in the platelet membranes from the propositus, half of the normal amount being present in both parents. These findings indicate CE to be a rapid, sensitive and reliable tool to investigate patients with abnormalities of the GP IIb-IIIa complex.

AB - Glanzmann thrombasthenia (GT) is an inherited hemorrhagic defect due to a failure of the platelet membrane glycoprotein (GP) IIb-IIIa complex. Capillary electrophoresis (CE) analysis of solubilized platelet membranes from normal individuals showed the presence of two peaks with a migration time of 27 and 29 min, respectively. An excellent run-to-run and day-to-day reproducibility of the technique (<1% variation of the retention time) was documented. Using an automated Ferguson method, the apparent molecular masses were 100.0 kDa and 138.5 kDa, respectively. Immunoprecipitation with monoclonal antibodies anti-GP IIIa (B59.2.1) and anti-IIb (61.9.1.3) showed the two peaks as IIIa and IIb, respectively. Electropherograms of a GT young man showed the lack of both peaks. Less than 50% of each peak was present in his parents. Polyacrylamide gel electrophoresis (PAGE), immunoblotting, and flow cytometry analyses showed that GP IIb and IIIa were undetectable in the platelet membranes from the propositus, half of the normal amount being present in both parents. These findings indicate CE to be a rapid, sensitive and reliable tool to investigate patients with abnormalities of the GP IIb-IIIa complex.

KW - Capillary electrophoresis

KW - Glanzmann's thrombasthenia

KW - Glycoproteins

KW - Platelet membrane

UR - http://www.scopus.com/inward/record.url?scp=0031806453&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031806453&partnerID=8YFLogxK

U2 - 10.1002/elps.1150190842

DO - 10.1002/elps.1150190842

M3 - Article

C2 - 9694297

AN - SCOPUS:0031806453

VL - 19

SP - 1468

EP - 1474

JO - Electrophoresis

JF - Electrophoresis

SN - 0173-0835

IS - 8-9

ER -