Novel monoclonal antibody recognizing triglyceride-rich oxidized LDLs associated with severe liver disease and small oxidized LDLs in normal subjects

Author:

Sakurai Toshihiro1,Ichikawa Ayako12,Furukawa Hiroyuki3,Wada Norio4,Nagasaka Atsushi4,Takahashi Yuji15,Fujikawa Masato5,Ikuta Akiko1,Furumaki Hiroaki1,Shiga Maiko2,Shimizu Chikara2,Hui Shu-Ping1,Jin Shigeki1,Takeda Seiji1,Fuda Hirotoshi1,Nagasaka Hironori6,Kobayashi Seiichi1,Chiba Hitoshi1

Affiliation:

1. Faculty of Health Sciences, Hokkaido University, Kita-12, Nishi-5, Kita-ku, Sapporo 060-0812

2. Division of Laboratory and Transfusion Medicine, Hokkaido University Hospital, Sapporo 060-8648

3. Department of Organ Transplantation and Regeneration, Hokkaido University Graduate School of Medicine, Sapporo 060-8638

4. Department of Internal Medicine

5. Department of Clinical Laboratory, Sapporo City General Hospital, Sapporo 060-8604

6. Department of Pediatrics, Takarazuka City Hospital, Takarazuka 665-0827, Japan

Abstract

Background Triglyceride-rich low-density lipoproteins (TG-rich LDLs) in the plasma of patients with severe liver disease are reported to change macrophages into foam cells in vitro. Methods Male BALB/c mice were immunized with TG-rich LDLs isolated from the plasma of a patient with severe liver disease. The resulting monoclonal antibody (G11-6) was used in a sandwich enzyme-linked immunosorbent assay (ELISA) in combination with polyclonal anti-apolipoprotein B antibodies. The time course of copper-mediated LDL oxidation was monitored using this ELISA. The results were compared with those of the two commercial ELISAs for oxidized LDLs using DLH or ML25, thiobarbituric acid reactive substances and the optical absorbance for the conjugated dienes generated in lipid peroxides. Furthermore, the lipoprotein fractions separated by gel filtration were tested with this ELISA in healthy volunteers ( n = 11) and patients ( n = 3) with liver disease. Results G11-6 reacted with oxidized LDLs during only the early phase of copper oxidation, being distinct from the other monoclonal antibodies and methods. G11-6 was confirmed to react with TG-rich LDLs in patients, while it reacted with small LDL particles in normal controls. Conclusions The monoclonal antibody G11-6 is useful for detecting oxidized small LDLs in normal controls and oxidized TG-rich LDLs in patients with severe liver disease.

Publisher

SAGE Publications

Subject

Clinical Biochemistry,General Medicine

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