Characterization of the epitopes specific for the monoclonal antibody 9F5-3a and quantification of oxidized HDL in human plasma

Ann Clin Biochem 2004;41:309-315
doi:10.1258/0004563041201491
© 2004 Royal Society of Medicine

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Original Articles


Takanori Nakajima,
Toshiyuki Matsunaga,
Shin-Ichiroh Kawai,
Shigeru Hokari,
Ikuo Inoue,
Shigehiro Katayama,
Atsuo Nagata and
Tsugikazu Komoda


Department of Biochemistry, Saitama Medical School, 38 Morohongo, Moroyama, Saitama 350-0495, Japan;
Department of Biochemistry, Saitama Medical School, 38 Morohongo, Moroyama, Saitama 350-0495, Japan;
Department of Biochemistry, Saitama Medical School, 38 Morohongo, Moroyama, Saitama 350-0495, Japan;
Department of Biochemistry, Saitama Medical School, 38 Morohongo, Moroyama, Saitama 350-0495, Japan;
Fourth Department of Internal Medicine, Saitama Medical School, 38 Morohongo, Moroyama, Saitama 350-0495, Japan;
Fourth Department of Internal Medicine, Saitama Medical School, 38 Morohongo, Moroyama, Saitama 350-0495, Japan;
Immunology Laboratory, Diagnostic Division, Yamasa Shoyu Co., Choshi, Chiba 288-0056, Japan;
Department of Biochemistry, Saitama Medical School, 38 Morohongo, Moroyama, Saitama 350-0495, Japan


Background: We previously isolated a monoclonal antibody, 9F5-3a, that is specific for HDL oxidized by CuSO4.

Methods: We examined the characteristics of the 9F5-3a epitopeby Western blot and measured the concentration of oxidized HDLin human plasma by enzyme-linked immunosorbent assay using thisantibody.

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) µg/L in 23 healthy controls, 191 (65) µg/L in 30 patients with non-insulin-dependent diabetes mellitus (P < 0.01 versus healthy controls) and 200 (87) µg/L in 25 patients with coronary artery disease (P < 0.01 versus healthycontrols). The concentrations of oxidized HDL did not correlatewith the concentrations of thiobarbituric acid-reactive substances.

Conclusions: The results indicate that determination of oxidizedHDL concentration may be useful for identifying patients withatherosclerotic disease.


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