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Journal of Materials Chemistry B Vol -1

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

Content Provider	Royal Society of Chemistry (RSC)
Author	Yamashiro-Kanashiro, E. H. G. Sales, M. Goreti F. Gidlund, Magnus Cabral-Miranda, Gustavo
Copyright Year	2014
Abstract	Increased levels of plasma oxLDL, which is the oxidized fraction of Low Density Lipoprotein (LDL), are associated with atherosclerosis, an inflammatory disease, and the subsequent development of severe cardiovascular diseases that are today a major cause of death in modern countries. It is therefore important to find a reliable and fast assay to determine oxLDL in serum. A new immunosensor employing three monoclonal antibodies (mAbs) against oxLDL is proposed in this work as a quick and effective way to monitor oxLDL. The oxLDL was first employed to produce anti-oxLDL monoclonal antibodies by hybridoma cells that were previously obtained. The immunosensor was set-up by self-assembling cysteamine (Cyst) on a gold (Au) layer (4 mm diameter) of a disposable screen-printed electrode. Three mAbs were allowed to react with N-hydroxysuccinimide (NHS) and ethyl(dimethylaminopropyl)carbodiimide (EDAC), and subsequently incubated in the Au/Cys. Albumin from bovine serum (BSA) was immobilized further to ensure that other molecules apart from oxLDL could not bind to the electrode surface. All steps were followed by various characterization techniques such as electrochemical impedance spectroscopy (EIS) and square wave voltammetry (SWV). The analytical operation of the immunosensor was obtained by incubating the sensing layer of the device in oxLDL for 15 minutes, prior to EIS and SWV. This was done by using standard oxLDL solutions prepared in foetal calf serum, in order to simulate patient's plasma with circulating oxLDL. A sensitive response was observed from 0.5 to 18.0 μg mL−1. The device was successfully applied to determine the oxLDL fraction in real serum, without prior dilution or necessary chemical treatment. The use of multiple monoclonal antibodies on a biosensing platform seemed to be a successful approach to produce a specific response towards a complex multi-analyte target, correlating well with the level of oxLDL within atherosclerosis disease, in a simple, fast and cheap way.
Starting Page	477
Ending Page	484
Page Count	8
File Format	HTM / HTML PDF
ISSN	2050750X
Volume Number	2
Issue Number	5
Journal	Journal of Materials Chemistry B
DOI	10.1039/c3tb21048k
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	NHS EDAC BSA EIS Low Density Lipoprotein SWV Blood plasma Lipoprotein Low-density lipoprotein Atherosclerosis Monoclonal antibody Hybridoma technology Cysteamine Cyst Electrode N-Hydroxysuccinimide Carbodiimide Albumin Electrochemistry Electrical impedance Spectroscopy Square wave Voltammetry
Content Type	Text
Resource Type	Article
Subject	Chemistry Medicine Materials Science Biomedical Engineering

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