Immunoreactivity of apolipoprotein B-100 in oxidatively modified low density lipoprotein.
Thirteen monoclonal antibodies (MAbs) against apolipoprotein B-100 (apo B) were used to analyze changes in immunoreactivity of human LDL resulting from oxidation mediated by cupric ions and oxygen. Decrease in immunoreactivity of oxidized LDL was demonstrated by competitive ELISA with MAbs 5F8, BL3, Mb43, 2G8, B3, B5, and BL7 for which the epitopes are located within residues 1-1297, 4235-4355, 4027-4081, 3728-4306, 2239-2331, 1854-1878, and in the vicinity of residue 2331, respectively. Immunoreactivity of the epitope B6 (2239-2331) increased during first 4 hours of oxidation and then diminished gradually. Epitope B1 (405-539) had slightly reduced immunoreactivity during first 8 h of LDL oxidation and then its minor increase was observed. MAb 12G10, specific to the epitope within apo B thrombin-digest fragment T4 (1-1297), displayed either weak or strong binding to LDL. LDL with weak binding pattern demonstrated significant increase in immunoreactivity upon oxidation. In contrast, LDL with strong binding pattern showed little to no change. Epitopes Mb47 (3441-3569) and 8G4 (1-1297) remained unchanged in oxidized LDL. Immunoreactivity of apo B-100 epitope recognized by MAb 4C11 (residues 2377-2658) was shown to be a function of oxidation time: it increased progressively up to 16 h and was stabilized for another 24 h of LDL oxidation. This epitope may be unmasked by LDL oxidation and may provide a useful immunochemical marker to monitor the extent of LDL oxidation.
Valentinova
N V
NV
Department of Medicine, Baylor College of Medicine, Houston, Texas 77030.
Gu
Z W
ZW
Yang
M
M
Yanushevskaya
E V
EV
Antonov
I V
IV
Guyton
J R
JR
Smith
C V
CV
Gotto
A M
AM
Yang
C Y
CY
D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D053299:Apolipoprotein B-100; D001055:Apolipoproteins B / Q000276:immunology*; D003300:Copper / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D006801:Humans; D008077:Lipoproteins, LDL / Q000737:chemistry / Q000276:immunology*; D010084:Oxidation-Reduction; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism
D000911:Antibodies, Monoclonal; D053299:Apolipoprotein B-100; D001055:Apolipoproteins B; D000939:Epitopes; D008077:Lipoproteins, LDL; D017392:Thiobarbituric Acid Reactive Substances; D003300:Copper