US 12,066,441 B2
Methods, reagents and kits for flow cytometric immunophenotyping
Jacobus Johannes Maria van Dongen, Rotterdam (NL); José Alberto Orfao de Matos Correia e Vale, Salamanca (ES); Juan Alejandro Flores-Montero, Salamanca (ES); Julia Maria Almeida Parra, Salamanca (ES); Vincent Henricus Johannes van der Velden, Capelle aan den IJssel (NL); Sebastian Böttcher, Stampe (DE); Andrew Craig Rawstron, Yorkshire (GB); Ruth Mary de Tute, West Yorkshire (GB); Ludovic Bernard Simon Lhermitte, Paris (FR); Vahid Asnafi, Bezons (FR); Ester Mejstríková, Prague (CZ); Tomasz Szczepanski, Zabrze (PL); Paulo Jorge Monteiro Da Silva Lucio, Carnaxide (PT); Marta Martin Ayuso, Salamanca (ES); and Carlos Eduardo Pedreira, Rio de Janeiro (BR)
Assigned to Erasmus University Medical Center Rotterdam, Rotterdam (NL)
Filed by Erasmus University Medical Center Rotterdam, Rotterdam (NL)
Filed on Mar. 8, 2021, as Appl. No. 17/195,416.
Application 17/195,416 is a continuation of application No. 15/849,196, filed on Dec. 20, 2017, granted, now 10,942,171.
Application 15/849,196 is a continuation of application No. 13/376,103, granted, now 9,880,158, previously published as PCT/NL2010/050332, filed on Jun. 2, 2010.
Claims priority of application No. 09161870 (EP), filed on Jun. 3, 2009.
Prior Publication US 2021/0263017 A1, Aug. 26, 2021
Int. Cl. G01N 33/533 (2006.01); G01N 15/14 (2006.01); G01N 33/49 (2006.01); G01N 33/50 (2006.01); G01N 33/532 (2006.01); G01N 33/569 (2006.01); G01N 15/1404 (2024.01)
CPC G01N 33/56972 (2013.01) [G01N 15/14 (2013.01); G01N 33/4915 (2013.01); G01N 33/5005 (2013.01); G01N 33/5091 (2013.01); G01N 33/5094 (2013.01); G01N 33/532 (2013.01); G01N 33/533 (2013.01); G01N 2015/1402 (2013.01); G01N 2015/1413 (2013.01)] 16 Claims
 
1. A reagent composition for flow cytometric immunophenotyping and characterization of myeloid cell subpopulations, wherein the reagent composition comprises antibodies against the markers human leukocyte antigen-DR (HLADR), CD45, CD34 and CD117, and wherein the reagent composition consists essentially of fluorochrome-conjugated antibodies that are directed against each marker within at least one of the following panels of markers:
(i) HLADR, CD45, CD16, CD13, CD34, CD117, CD11b and CD10, wherein the antibody against HLADR, the antibody against CD45, the antibody against CD16, the antibody against CD13, the antibody against CD34, the antibody against CD117, the antibody against CD11b, and the antibody against CD10 are each conjugated to a fluorochrome that has a different fluorescence emission from the other fluorochromes;
(ii) HLADR, CD45, CD35, CD64, CD34, CD117, immune receptor expressed by myeloid cells-2 (IREM2) and CD14, wherein the antibody against HLADR, the antibody against CD45, the antibody against CD35, the antibody against CD64, the antibody against CD34, the antibody against CD117, the antibody against IREM2, and the antibody against CD14 are each conjugated to a fluorochrome that has a different fluorescence emission from the other fluorochromes;
(iii) HLADR, CD45, CD36, CD105, CD34, CD117, CD33 and CD71, wherein the antibody against HLADR, the antibody against CD45, the antibody against CD36, the antibody against CD105, the antibody against CD34, the antibody against CD117, the antibody against CD33, and the antibody against CD71 are each conjugated to a fluorochrome that has a different fluorescence emission from the other fluorochromes;
(iv) HLADR, CD45, CD15, neuron-glial antigen-2 (NG2), CD34, CD117, CD7 and CD38, wherein the antibody against HLADR, the antibody against CD45, the antibody against CD15, the antibody against NG2, the antibody against CD34, the antibody against CD117, the antibody against CD7, and the antibody against CD38 are each conjugated to a fluorochrome that has a different fluorescence emission from the other fluorochromes;
(v) HLADR, CD45, CD42a, CD61, CD203c, CD34, CD117, CD123 and CD4, wherein the antibodies against CD42a and CD61 are conjugated to the same fluorochrome, and wherein the antibody against HLADR, the antibody against CD45, the antibody against CD203c, the antibody against CD34, the antibody against CD117, and the antibody against CD4 are each conjugated to a fluorochrome that has a different fluorescence emission from the other fluorochromes;
(vi) HLADR, CD45, CD41, CD25, CD34, CD117, CD42b and CD9, wherein the antibody against HLADR, the antibody against CD45, the antibody against CD41, the antibody against CD25, the antibody against CD34, the antibody against CD117, the antibody against CD42b, and the antibody against CD9 are each conjugated to a fluorochrome that has a different fluorescence emission from the other fluorochromes; or
(vii) HLADR, CD45, Terminal deoxynucleotidyl transferase (TdT), CD56, CD34, CD117, CD22 and CD19 wherein the antibody against HLADR, the antibody against CD45, the antibody against TdT, the antibody against CD56, the antibody against CD34, the antibody against CD117, the antibody against CD22, and the antibody against CD19 are each conjugated to a fluorochrome that has a different fluorescence emission from the other fluorochromes.