Abstract
This chapter presents a study on the humoral immune response to oxidized low-density lipoprotein (oxi-LDL) in patients with coronary artery disease (CAD). Blood serum samples were collected by vein puncture of 51 patients with angiographically proven CAD and 51 age-matched controls. LDL was purified from patient's blood plasma by density gradient centrifugation and oxidized by treatment with peroxidase. Both native low-density lipoprotein (nLDL) and oxi-LDL were used for development of an immunoassay for detection of autoantibodies against oxidized LDL in patients with coronary artery disease. To demonstrate the specific character of this immunoassay, serial dilutions of positively reacting human serum were tested against a fixed amount of oxi-LDL coated on PVC wells. Peripheral blood leucocytes (PBL) from eight patients with CAD or insulin-dependent diabetes mellitus (IDDM) were used in independent fusion experiments to produce heterohybridomas secreting monoclonal antibodies against oxi-LDL. Assessment of the specificity of each monoclonal antibody demonstrated that the heterohybridoma designated as IvNi 2D2 secreted an IgG antibody that reacted with oxi-LDL in enzyme-linked immunoassay (ELISA).
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