Flow cytometry and immunoblotting analysis of monoclonal antibodies directed to complement regulatory proteins.
A series of 18 monoclonal antibodies directed to complement regulatory proteins were investigated by flow cytometry and immunoblotting. Seventeen antibodies are directed against a phosphatidyl inositol-linked glycoprotein since they show a dual population of erythrocytes from a paroxysmal nocturnal haemoglobinuria (PNH) patient. From this group, 6 antibodies revealed a 65-70 kDa band on immunoblots allowing their identification as anti-DAF (Decay Accelerating Factor; CD55 antigen), and 11 bound to a 20 kDa molecule corresponding to MIRL (Membrane Inhibitor of Reactive lysis, CD59 antigen). One antibody revealed an homogeneous population from the PNH patient and bound to a 200 kDa band on immunoblot that might corresponds to the CR1 (Complement Receptor type 1; CD35).
Blanchard
D
D
Etablissement de Transfusion Sanguine de Loire Atlantique/Vendée, Site de Nantes, France.
Navenot
J M
JM
Petit-Le Roux
Y
Y
Willem
C
C
Loirat
M J
MJ
D000911:Antibodies, Monoclonal; D018958:CD55 Antigens / Q000276:immunology*; D018957:CD59 Antigens / Q000276:immunology*; D016022:Case-Control Studies; D004912:Erythrocytes / Q000276:immunology*; D005434:Flow Cytometry; D006457:Hemoglobinuria, Paroxysmal / Q000097:blood / Q000276:immunology*; D006801:Humans; D015151:Immunoblotting; D017463:Receptors, Complement 3b / Q000276:immunology*
D000911:Antibodies, Monoclonal; D018958:CD55 Antigens; D018957:CD59 Antigens; D017463:Receptors, Complement 3b