RESUMO
This paper presents a hand-held point-of-care device that incorporates a lab-on-a-chip module with interdigitated capacitive biosensors for label-free detection of multiple cancer and cardiovascular disease biomarkers. The developed prototype is comprised of a cartridge incorporating capacitive biodetection sensors, a sensitive capacitive readout electronics enclosed in a hand-held unit, and data analysis software calculating the concentration of biomarkers using previously stored reference database. The capacitive biodetection sensors are made of interdigitated circular electrodes, which are preactivated with single (for detecting one biomarker) or multiple specific antibodies (for detecting multiple disease biomarkers). Detection principle of capacitive biosensor is based on measuring the level of capacitance change between interdigitated electrode pairs induced by the change in dielectric constant due to affinity-based electron exchange in between antibodies/antigens and electrodes. The more antibody-antigens binding occurs, the more capacitance change is measured due to the change in dielectric constant of the capacitance media. The device uses preactivated ready-to-use cartridges embedded with capacitive biosensors with shelf-life of three months under optimal conditions, and is capable of onsite diagnosis and can report the result in less than 30 min. The device is verified with real patient blood samples for six different disease biomarkers.
Assuntos
Biomarcadores/sangue , Técnicas Biossensoriais/instrumentação , Cardiopatias/diagnóstico , Neoplasias/diagnóstico , Sistemas Automatizados de Assistência Junto ao Leito , Biomarcadores Tumorais/sangue , Bases de Dados Factuais , Desenho de Equipamento , Humanos , Dispositivos Lab-On-A-ChipRESUMO
The detection and quantification of cancer biomarkers in human blood is crucial to diagnose patients in the early stage of a disease. The recent advances in biosensor technology can improve detection by reducing the application time and cost without an invasive approach. In this study, a highly sensitive, novel nanoparticle-modified capacitive sensor was developed for the detection of cancer markers. The current work mainly focused on developing a surface modification protocol for achieving higher sensitivity using Au-NPs. An interdigitated electrode (IDE) transducer was modified using gold nanoparticles (Au-NPs) for signal enhancement, the platform was initially optimized with a small size IL-6 protein and the methodology was then applied for multiple marker detection with the aim of precise disease diagnostics. Carcinoembryonic antigen (CEA) and epidermal growth factor receptor (hEGFR) could be successfully detected in the concentration range of 20-1000 pg mL(-1) while cancer antigen 15-3 (CA15-3) was detected in the range of 10-200 U mL(-1). These results show an increase of sensitivity by five-fold with respect to those not modified, demonstrating a highly sensitive and specific capacitive immunoassay that has a great potential for the use of early diagnosis of cancer disease.
Assuntos
Biomarcadores Tumorais/sangue , Técnicas Biossensoriais/métodos , Eletrodos , Ouro/química , Imunoensaio/métodos , Neoplasias Pulmonares/diagnóstico , Nanopartículas Metálicas/química , Antígeno Carcinoembrionário/sangue , Receptores ErbB/sangue , Humanos , Neoplasias Pulmonares/sangue , Mucina-1/sangueRESUMO
The immobilisation of biological recognition elements onto a sensor chip surface is a crucial step for the construction of biosensors. While some of the optical biosensors utilise silicon dioxide as the sensor surface, most of the biosensor surfaces are coated with metals for transduction of the signal. Biological recognition elements such as proteins can be adsorbed spontaneously on metal or silicon dioxide substrates but this may denature the molecule and can result in either activity reduction or loss. Self assembled monolayers (SAMs) provide an effective method to protect the biological recognition elements from the sensor surface, thereby providing ligand immobilisation that enables the repeated binding and regeneration cycles to be performed without losing the immobilised ligand, as well as additionally helping to minimise non-specific adsorption. Therefore, in this study different surface chemistries were constructed on SPR sensor chips to investigate protein and DNA immobilisation on Au surfaces. A cysteamine surface and 1%, 10% and 100% mercaptoundecanoic acid (MUDA) coatings with or without dendrimer modification were utilised to construct the various sensor surfaces used in this investigation. A higher response was obtained for NeutrAvidin immobilisation on dendrimer modified surfaces compared to MUDA and cysteamine layers, however, protein or DNA capture responses on the immobilised NeutrAvidin did not show a similar higher response when dendrimer modified surfaces were used.
Assuntos
DNA/análise , Proteínas/análise , Ressonância de Plasmônio de Superfície , Técnicas Biossensoriais , Cisteamina/química , Dendrímeros/química , Ouro/química , Hibridização de Ácido Nucleico , Propriedades de SuperfícieRESUMO
An immunoassay in optimised conditions with a highly sensitive surface plasmon resonance (SPR) based biosensor was developed for the detection of the cancer biomarker carcinoembryonic antigen (CEA). Different formats of the immunoassay were initially investigated on the surface of the gold sensor chip. A self-assembled monolayer (SAM) was formed on the gold chip using 11-mercaptoundecanoic acid (MUDA), before the immobilisation of the antibodies was conducted. The assay was then formed in a direct capture and a sandwich assay. In order to increase the sensor signal the CEA antigen was incubated with the detection/capture antibody before it was injected to the sensor chip surface and the results were recorded in real-time using the Biacore 3000 instrument. A detection limit of 3 ng ml(-1) CEA was obtained with a dynamic detection range from 3 ng ml(-1) to 400 ng ml(-1) with correlation coefficients of 1.00 and 0.99 for the sandwich and rabbit anti-mouse (RAM) capture assay. Kinetic data analysis was performed for the standard capture test and subsequently for the developed assays and R(max) showed an increase from 215 RU for the standard capture test to 428 RU for the RAM-capture assay and 734 RU for the sandwich assay, respectively. The developed SPR immunosensor using the sandwich assay format showed high sensitivity and reproducibility for CEA detection which makes it a promising procedure for cancer biomarker analysis.