RESUMO
A sandwich-type nanostructured immunosensor based on carboxylated multi-walled carbon nanotube (CMWCNT)-embedded whiskered nanofibres (WNFs) was developed for detection of cardiac Troponin I (cTnI). WNFs were directly fabricated on glassy carbon electrodes (GCE) by removing the sacrificial component (polyethylene glycol, PEG) after electrospinning of polystyrene/CMWCNT/PEG nanocomposite nanofibres, and utilised as a transducer layer for enzyme-labeled amperometric immunoassay of cTnI. The whiskered segments of CMWCNTs were activated and utilised to immobilise anti-cTnT antibodies. It was observed that the anchored CMWCNTs within the nanofibres were suitably stabilised with excellent electrochemical repeatability. A sandwich-type immuno-complex was formed between cTnI and horseradish peroxidase-conjugated anti-cTnI (HRP-anti-cTnI). The amperometric responses of the immunosensor were studied using cyclic voltammetry (CV) through an enzymatic reaction between hydrogen peroxide and HRP conjugated to the secondary antibody. The nanostructured immunosensor delivered a wide detection range for cTnI from the clinical borderline for a normal person (0.5-2ngmL-1) to the concentration present in myocardial infarction patients (> 20ngmL-1), with a detection limit of ~ 0.04ngmL-1. It also showed good reproducibility and repeatability for three different cTnI concentration (1, 10 and 25ngmL-1) with satisfactory relative standard deviations (RSD). Hence, the proposed nanostructured immunosensor shows potential for point-of-care testing.
Assuntos
Técnicas Biossensoriais , Técnicas Eletroquímicas , Imunoensaio , Nanofibras/química , Nanotubos de Carbono/química , Troponina I/sangue , Anticorpos Imobilizados/química , Anticorpos Monoclonais/química , Eletrodos , Peroxidase do Rábano Silvestre/química , Humanos , Peróxido de Hidrogênio/química , Imunoconjugados/química , Limite de Detecção , Nanocompostos/química , Nanocompostos/ultraestrutura , Nanofibras/ultraestrutura , Nanotubos de Carbono/ultraestrutura , Polietilenoglicóis/química , Polietilenoglicóis/isolamento & purificação , Poliestirenos/química , Reprodutibilidade dos TestesRESUMO
The vital importance of early and accurate diagnosis of cardiovascular diseases (CVDs) to prevent the irreversible damage or even death of patients has driven the development of biosensor devices for detection and quantification of cardiac biomarkers. Electrochemical biosensors offer rapid sensing, low cost, portability and ease of use. Over the past few years, nanotechnology has contributed to a tremendous improvement in the sensitivity of biosensors. In this review, the authors summarise the state-of-the-art of the application of one particular type of nanostructured material, i.e. nanofibres, for use in electrochemical biosensors for the ultrasensitive detection of cardiac biomarkers. A new way of classifying the nanofibre-based electrochemical biosensors according to the electrical conductance and the type of nanofibres is presented. Some key data from each article reviewed are highlighted, including the mechanism of detection, experimental conditions and the response range of the biosensor. The primary aim of this review is to emphasise the prospects for nanofibres for the future development of biosensors in diagnosis of CVDs as well as considering how to improve their characteristics for application in medicine.
