TY - JOUR
T1 - Immunologic detection and measurement of hypochlorite-modified LDL with specific monoclonal antibodies
AU - Malle, Ernst
AU - Hazell, Linda
AU - Stocker, Roland
AU - Sattler, Wolfgang
AU - Esterbauer, Hermann
AU - Waeg, Georg
N1 - Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.
PY - 1995/7
Y1 - 1995/7
N2 - Oxidation of LDL is thought to contribute to the early stages of atherogenesis. Because myeloperoxidase is present in atherosclerotic lesions and can produce the strong oxidant hypochlorous acid (HOCl), which converts LDL into its high-uptake atherogenic form in vitro, we raised polyclonal and monoclonal antibodies (MoAbs) against HOCl-modified LDL (HOCl-LDL). Characterization of the polyclonal antihuman HOCl-LDL Abs showed that they cross-reacted strongly with 4-hydroxynomenal-malondialdehyde-, and Cu2+oxidized LDL. Similarly, polyclonal and some monoclonal Abs against aldehyde- anti Cu2+-modified LDL cross-reacted with HOCl-LDL. In contrast to the polyclonal Abs, two selected hybridoma cell line supernatants containing MoAbs raised against HOCl-LDL (MoAb-A and MoAb-B) did not cross- react with either native LDL or aldehyde or Cu2+-modified LDL. MoAb-A (clone IB10A11, subtype IgG1κ) recognized an epitope that appeared to be specific for HOCl-LDL and depended on the tertiary structure of the (lipo)protein, as judged by a lack of cross-reactivity with HOCl-modified human and bovine serum albumin and a loss of reactivity associated with lipoprotein denaturation. MoAb-B (clone 2D10G9, subtype lgG2bκ), on the other hand, gave identical titration curves with HOCl-LDL and HOCl-modified albumins, suggesting that this antibody recognized epitopes that are commonly generated on proteins that have been oxidized with HOCl. Thus, MoAb-A and MoAb-B may be useful tools for the investigation of a possible role for HOCl- mediated damage to (lipo)proteins in atherosclerosis and other inflammatory diseases.
AB - Oxidation of LDL is thought to contribute to the early stages of atherogenesis. Because myeloperoxidase is present in atherosclerotic lesions and can produce the strong oxidant hypochlorous acid (HOCl), which converts LDL into its high-uptake atherogenic form in vitro, we raised polyclonal and monoclonal antibodies (MoAbs) against HOCl-modified LDL (HOCl-LDL). Characterization of the polyclonal antihuman HOCl-LDL Abs showed that they cross-reacted strongly with 4-hydroxynomenal-malondialdehyde-, and Cu2+oxidized LDL. Similarly, polyclonal and some monoclonal Abs against aldehyde- anti Cu2+-modified LDL cross-reacted with HOCl-LDL. In contrast to the polyclonal Abs, two selected hybridoma cell line supernatants containing MoAbs raised against HOCl-LDL (MoAb-A and MoAb-B) did not cross- react with either native LDL or aldehyde or Cu2+-modified LDL. MoAb-A (clone IB10A11, subtype IgG1κ) recognized an epitope that appeared to be specific for HOCl-LDL and depended on the tertiary structure of the (lipo)protein, as judged by a lack of cross-reactivity with HOCl-modified human and bovine serum albumin and a loss of reactivity associated with lipoprotein denaturation. MoAb-B (clone 2D10G9, subtype lgG2bκ), on the other hand, gave identical titration curves with HOCl-LDL and HOCl-modified albumins, suggesting that this antibody recognized epitopes that are commonly generated on proteins that have been oxidized with HOCl. Thus, MoAb-A and MoAb-B may be useful tools for the investigation of a possible role for HOCl- mediated damage to (lipo)proteins in atherosclerosis and other inflammatory diseases.
KW - atherosclerosis
KW - lipid peroxidation
KW - myeloperoxidase
KW - oxidized lipoproteins
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U2 - 10.1161/01.ATV.15.7.982
DO - 10.1161/01.ATV.15.7.982
M3 - Article
C2 - 7541296
AN - SCOPUS:0029036325
SN - 1079-5642
VL - 15
SP - 982
EP - 989
JO - Arteriosclerosis, Thrombosis, and Vascular Biology
JF - Arteriosclerosis, Thrombosis, and Vascular Biology
IS - 7
ER -