Immunological detection of large oxidized lipoproteins in hypertriglyceridemic serum

Abstract

Background: Triglyceride-rich, low-density lipoproteins (TG-rich LDL) have been reported as an oxidized lipoprotein species in patients with severe liver disease. Using TG-rich LDL as an immunogen, we obtained a monoclonal antibody (G11-6) that reacted with TG-rich LDL from patients with liver disease and with metal-oxidized LDL only in the early process of the oxidation reaction. This study determined the G11-6-reactive lipoproteins in hypertriglyceridemic serum. Methods: Serum samples from healthy volunteers (n ¼ 12) and hypertriglyceridemic patients (n ¼ 9) were fractionated by gel filtration and subjected to a sandwich enzyme-linked immunosorbent assay (ELISA) using G11-6 and polyclonal anti-apolipoprotein B antibodies. Results: Small LDL and larger lipoproteins reacted with G11-6. G11-6-reactive small LDL was identified in both the healthy subjects and hypertriglyceridemic patients, whereas G11-6-reactive larger lipoproteins were found only in the hypertriglyceridemic patients. Conclusions: G11-6 is a useful tool for detecting increased large oxidized lipoproteins in hypertriglyceridemic patients.

Specific label-free and real-time detection of oxidized low density lipoprotein (oxLDL) using an immunosensor with three monoclonal antibodies

“…There are many ways to determine oxLDL in the blood by ELISA (enzyme-linked immunosorbent assay) methods, such as employing different monoclonal and polyclonal antibodies that are capable of detecting oxLDL fractions involved in the atherogenesis. [11][12][13] Alternative methods employ anti-oxLDL Abs conjugated to gold nanoparticles and subsequent detection by liquid chromatography/tandem-mass spectrometry (LC-MS/MS) 13 or proteoheparan sulfate coated hydrophobic silica for subsequent ellipsometry readings. [14][15][16] These two approaches are however unsuitable for applications in point-of-care diagnosis, also being coupled to expensive equipment.…”

Novos biossensores baseados em anticorpos naturais e sintéticos para detecção de LDL oxidada (oxLDL) usada como biomarcador de aterosclerose.

“…Atualmente existem várias formas de determinar oxLDL no sangue pelo método de ELISA empregando anticorpos monoclonais e policlonais, capazes de detectar pequenas frações de oxLDL (ITABE; SAKURAI et al, 2013). Apesar de haver uma ampla diferença nos procedimentos de ELISA utilizados para determinar oxLDL, desde as partículas alvo para serem detectadas, até a avidez que os métodos proporcionam, não há muita mudança nos procedimentos que levem a uma melhor aplicabilidade na rotina para diagnóstico.…”