Premium
Relative quantification of beta‐adrenergic receptor in peripheral blood cells using flow cytometry
Author(s) -
Saygin Didem,
Wanner Nicholas,
Rose Jonathan A.,
Naga Prasad Sathyamangla V.,
Tang W. H. Wilson,
Erzurum Serpil,
Asosingh Kewal
Publication year - 2018
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.23358
Subject(s) - flow cytometry , biotinylation , receptor , alprenolol , cytometry , microbiology and biotechnology , chemistry , adrenergic receptor , biology , biochemistry
Beta‐adrenergic receptors (β‐ARs) play a critical role in many diseases. Quantification of β‐AR density may have clinical implications in terms of assessing disease severity and identifying patients who could potentially benefit from beta‐blocker therapy. Classical methods for β‐AR quantification are based on labor‐intensive and time‐consuming radioligand binding assays. Here, we report optimization of a flow cytometry‐based method utilizing a biotinylated β‐AR ligand alprenolol as a probe and use of this method to quantify relative receptor expression in healthy controls (HC). Quantum™ MESF beads were used for quantification in absolute fluorescence units. The probe was chemically modified by adding a spacer moiety between biotin and alprenolol to stabilize receptor binding, thus preventing binding decay. Testing of three different standard cell fixation and permeabilization methods (formaldehyde fixation and saponin, Tween‐20, or Triton‐X 100 permeabilization) showed that the formaldehyde/Triton‐X 100 method yielded the best results. β‐AR expression was significantly higher in granulocytes compared to mononuclear cells. These data show that flow cytometric quantification of relative β‐AR expression in circulating leukocytes is a suitable technology for large‐scale clinical application. © 2018 International Society for Advancement of Cytometry