Characterization of a specific monoclonal antibody 9F5-3a and the development of assay system for oxidized HDL.
We obtained a monoclonal antibody 9F5-3a against oxidative low-density lipoprotein (LDL) modified with CuSO4 and established a sandwich ELISA for detection of oxidized high-density lipoprotein (oxHDL). The 9F5-3a was reacted strongly with oxHDL and to a lesser degree with oxLDL and LDL. In contrast, little or no reactivity was found with HDL. When the generation of oxHDL was limited by the addition of alpha-tocopherol and catalase, reactivity to 9F5-3a was reduced. Incubation of oxHDL with excess lyso-phosphatidylcholine (lyso-PC) also reduced immunoreactivity, but not by only lyso-PC. These results suggested that the epitope is possibly associated with oxHDL-linked lyso-PC induced mainly by the hydroxyl radical.
Nakajima
T
T
1st Department of Biochemistry, Saitama Medical School, Japan.
Sakagishi
Y
Y
Katahira
T
T
Nagata
A
A
Kuwae
T
T
Nakamura
H
H
Inoue
I
I
Takahashi
K
K
Katayama
S
S
Komoda
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001161:Arteriosclerosis / Q000209:etiology; D003300:Copper / Q000737:chemistry; D019327:Copper Sulfate; D008075:Lipoproteins, HDL / Q000737:chemistry / Q000276:immunology*; D008077:Lipoproteins, LDL / Q000737:chemistry / Q000276:immunology*; D008244:Lysophosphatidylcholines / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D010084:Oxidation-Reduction
D000911:Antibodies, Monoclonal; D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL; D008244:Lysophosphatidylcholines; D003300:Copper; D019327:Copper Sulfate