Abstract
RBC flow cytometric analysis is usually used to quantify antigen content. Calibration systems enable antigen content determination by relating mean fluorescence intensity with the number of bound antibody molecules (equivalent to the number of antigen molecules). For that reason, antibodies must be used at saturating concentration, which may lead to agglutination when working with high density antigens. Then, forward scattering, side scattering and fluorescence will be increased, thus obtaining wrong results. In this work, the simple Langmuir adhesion model was applied. Flow cytometry was used to quantify GPA, a transmembrane protein present at high density on RBC. The fluorescence intensity of samples at different anti-GPA sub-saturating concentrations was measured. Sometimes, agglutinates were present and two peaks of fluorescence were observed, the principal one corresponding to isolated cells and the secondary one corresponding to agglutinated cells. In those cases, the principal peak was taken into account for the analysis. The GPA antigen content obtained for nine analyzed samples ranged from 3 to 13 x 10(5) sites per cell, which is similar to those values found in literature. Therefore, the Langmuir adsorption model enab...Continue Reading
References
Apr 1, 1992·Transfusion Medicine Reviews·J P Cartron, C Rahuel
Jan 1, 1991·Vox Sanguinis·Z N BernemanM E Peetermans
Jan 1, 1986·The Biochemical Journal·A H MerryD J Anstee
Feb 13, 1995·Journal of Immunological Methods·H JouinP Dubois
Feb 1, 1996·Cytometry·V L FerrandM J Bourdeaux
Jan 1, 1996·Transfusion clinique et biologique : journal de la Société française de transfusion sanguine·A FletcherM Stavliotis
Jan 1, 1996·Transfusion clinique et biologique : journal de la Société française de transfusion sanguine·W A Flegel, F F Wagner
Oct 17, 1998·Cytometry·S SerkeD Huhn
Oct 17, 1998·Cytometry·R LenkeiG Marti
Mar 16, 2000·Vox Sanguinis·G LarsonL Rydberg
Dec 21, 2000·British Journal of Haematology·M J KingK Chambers
Aug 8, 2001·Clinical and Laboratory Haematology·U OelschlaegelG Ehninger
Dec 26, 2001·Cytometry·D BratosinJ Montreuil
Mar 14, 2002·Transfusion clinique et biologique : journal de la Société française de transfusion sanguine·W A FlegelF F Wagner
Jun 27, 2002·Transfusion·Rainer LynenWolfgang R Mayr
Sep 5, 2002·Cytometry·J PichlerG Fritsch
Citations
Jul 19, 2013·PloS One·Santosh K MisraSantanu Bhattacharya
Sep 12, 2008·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·María Belén Jiménez-DíazIñigo Angulo-Barturen
Feb 8, 2011·Journal of Immunological Methods·Brian T Grimberg
Nov 15, 2011·Biochimica Et Biophysica Acta·S De OliveiraC Saldanha
Dec 11, 2007·Langmuir : the ACS Journal of Surfaces and Colloids·Darby KozakMatt Trau