United Kingdom (UK)
Recombinant monoclonal antibody to Oxidized phospholipid. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma E06.
Alternative Name(s) of Target: OxPL; OxLDL; CuOx-LDL; copper-oxidized low-density lipoproteins; EO6
Immunogen: The original mouse antibody was raised from apoE-deficient mice without any external immunogens. They were fed a high fat diet containing 12.8% milk fat and 0.15% cholesterol for 7 months. Such dietary intervention induces extensive aortic atherosclerosis, and is accompanied by the formation of high titers of E0 antibodies to MDA-lysine.
Specificity: The antibody is specific for oxidized lipoproteins in mice.
Application Notes: To characterize the mouse version of this antibody an antibody absorption experiment was performed on increasing concentrations of native PAPC, OxPAPC, or microemulsions made from lipids extracted from native LDL or CuOx-LDL. Further an ELISA was done on antigens in PBS using the mouse version of this antibody. The alkaline phosphatase–labeled goat anti–mouse IgM version of this antibodywas used to detect CuOx-LDL and native -LDL in a western blot assay. Also a flow cytometric experiment was performed on resident peritoneal macrophages isolated from female Swiss-Webster mice using the original mouse version of this antibody. And finally a binding and degradation assay was performed on mouse peritoneal macrophages using the mouse version of this antibody (Hörkkö et al, 1999; pmid:9884341). To identify whether the antibody binds to native or modified human LDL a radioimmunoassay was performed using the mouse version of this antibody. Immunocytochemistry was performed on atherosclerotic lesions of LDL receptor–deficient rabbits, balloon-catheterized, cholesterol-fed New Zealand white (NZW) rabbits, and human brain arteries using the mouse version of this antibody (Palinski et al, 1996; pmid:8698873). The extent of LDL oxidation Induced by copper was assessed using the mouse version of this antibody. This was done by a luminescence signal (relative light units) generated from streptavidin, which was conjugated to Aqualite bound to biotinylated antibodies (Sundell et al, 2003; pmid:12626663). To assess the binding profile of the scFv version of this antibody an ELISA was performed on phosphocholine epitopes as expressed on Cu-OxLDL, phosphocholine-KLH, PC-BSA, POVPC-BSA, and capsular polysaccharide (C-PS) of Streptococcus pneumonia, as well as to the anti-T15 idiotype antibody AB1-2 (Que et al, 29875409). The ability of the scFv version of this antibody to bind to apoptic cells was tested by flow cytometry on thymocytes collected from C57BL/6 mice induced to undergo apoptosis (Que et al, 29875409).
Antibody first published in:
Palinski et al. Cloning of monoclonal autoantibodies to epitopes of oxidized lipoproteins from apolipoprotein E-deficient mice. Demonstration of epitopes of oxidized low density lipoprotein in human plasma J Clin Invest. 1996 Aug 1;98(3):800-14. PMID:8698873
Note on publication:
Autoantibodies recognizing model epitopes of oxidized low density lipoprotein, such as malondialdehydelysine, occur in plasma and in atherosclerotic lesions of humans and animals. Because apo E-deficient mice develop particularly high titers of such autoantibodies, their spleens were used to clone 13 monoclonal antibodies to various epitopes of oxidized LDL ("E0 antibodies").