Characterization of a monoclonal antibody that binds to apolipoprotein E and to lipoprotein of human plasma containing apo E. Applications to ELISA quantification of plasma apo E.
This study describes the development of an enzyme-linked immunosorbent assay for human apolipoprotein E (apo E). A mouse monoclonal IgG1 antibody named E01 against apolipoprotein E was selected from five antibodies secreted by hybridomas. This antibody had a high affinity for apo E [K = 1.2 x 10(7) L.M-1 for purified apo E and K = 1.05 x 10(7) L.M-1 for native apo E in very low density lipoproteins) in liquid phase and recognized every isoform of apo E but not other proteins in VLDL. Competition experiments with 125I apo E showed that its binding affinity for the E in every density class (VLDL, HDL, LDL) and in serum was the same. This antibody was used for the quantification of human apo E in serum by enzyme linked immunoassay. E01 was coated on microtiter plates and a polyclonal peroxidase-conjugate was used as second antibody. A good correlation was observed between the values obtained for apo E using both monoclonal and polyclonal antibodies.
Leroy
A
A
SERLIA, Institut Pasteur, Lille, France.
Vu-dac
N
N
Koffigan
M
M
Clavey
V
V
Fruchart
J C
JC
D000818:Animals; D000911:Antibodies, Monoclonal; D000915:Antibody Affinity; D000918:Antibody Specificity; D001057:Apolipoproteins E / Q000276:immunology*; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008079:Lipoproteins, VLDL / Q000032:analysis; D051379:Mice
D000911:Antibodies, Monoclonal; D001057:Apolipoproteins E; D008079:Lipoproteins, VLDL