Characterization of the epitopes specific for the monoclonal antibody 9F5-3a and quantification of oxidized HDL in human plasma.
BACKGROUND
We previously isolated a monoclonal antibody, 9F5-3a, that is specific for HDL oxidized by CuSO(4).


METHODS
We examined the characteristics of the 9F5-3a epitope by Western blot and measured the concentration of oxidized HDL in human plasma by enzyme-linked immunosorbent assay using this antibody.


RESULTS
The monoclonal antibody specifically reacted with oxidized HDL in a mixture of HDL, LDL and modified lipoproteins. Oxidation of the HDL particles accelerated cross-linkage of apolipoproteins caused by lipid peroxidation, and the cross-linked apolipoprotein AI selectively reacted with the 9F5-3a antibody. Mean (standard deviation) plasma concentrations of oxidized HDL were 127 (50) microg/L in 23 healthy controls, 191 (65) microg/L in 30 patients with non-insulin-dependent diabetes mellitus (P < 0.01 versus healthy controls) and 200 (87) microg/L in 25 patients with coronary artery disease (P < 0.01 versus healthy controls). The concentrations of oxidized HDL did not correlate with the concentrations of thiobarbituric acid-reactive substances.


CONCLUSIONS
The results indicate that determination of oxidized HDL concentration may be useful for identifying patients with atherosclerotic disease.
Department of Biochemistry, Saitama Medical School, 38 Morohongo, Moroyama, Saitama 350-0495, Japan.
Takanori
T
Nakajima
Toshiyuki
T
Matsunaga
Shin-Ichiroh
S
Kawai
Shigeru
S
Hokari
Ikuo
I
Inoue
Shigehiro
S
Katayama
Atsuo
A
Nagata
Tsugikazu
T
Komoda
D000328:Adult; D000368:Aged; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D016022:Case-Control Studies; D003324:Coronary Artery Disease / Q000097:blood; D003924:Diabetes Mellitus, Type 2 / Q000097:blood; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry; D005260:Female; D006801:Humans; D008075:Lipoproteins, HDL / Q000097:blood* / Q000737:chemistry / Q000276:immunology*; D008297:Male; D008875:Middle Aged; D010084:Oxidation-Reduction
D000911:Antibodies, Monoclonal; D000939:Epitopes; D008075:Lipoproteins, HDL