A simultaneous negative and positive selection method that can detect chimerism at a frequency of 1 per 10,000 by flow cytometry.
A three-color flow cytometric method with a simultaneous negative and positive selection (SNAPS) approach has been developed to detect blood chimerism using a FITC-conjugated recipient-specific HLA monoclonal antibody, a biotinylated donor-specific HLA monoclonal antibody and Cy5-PE-conjugated streptavidin and a PE-conjugated CD3 monoclonal antibody. Detection of simulated blood chimerism at a frequency of 0.01% was achieved by SNAPS. In contrast, by one-color flow cytometry methods, chimerism can be detected at only about 3-4% with the same donor-specific HLA monoclonal antibody. The percentages of donor CD3 cells detected by SNAPS are proportional to the percentages of donor CD3 cells counted. SNAPS method provides a fast, sensitive and reliable way to analyze trace amounts of donor cells in a recipient's blood.
Pei
R
R
Department of Research, One Lambda Inc., Canoga Park, California, USA. rpei@onelambda.com
Chen
T
T
Orpilla
J
J
Lee
J H
JH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D001782:Blood Donors; D017252:CD3 Complex / Q000276:immunology; D002678:Chimera / Q000276:immunology*; D005434:Flow Cytometry / Q000379:methods*; D006680:HLA Antigens / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D007075:Immunoglobulin M / Q000032:analysis; D017404:In Situ Hybridization, Fluorescence; D008214:Lymphocytes / Q000276:immunology*; D051379:Mice; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D017252:CD3 Complex; D006680:HLA Antigens; D007074:Immunoglobulin G; D007075:Immunoglobulin M