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1.
Analyst ; 143(22): 5505-5511, 2018 Nov 05.
Artículo en Inglés | MEDLINE | ID: mdl-30295303

RESUMEN

We report a low-cost, sensitive, bead-based electrochemical immunoarray for soluble L-selectin (or CD62L protein), a potential biomarker for staging bladder cancer. We used a semi-automated modular microfluidic array with online antigen capture on superparamagnetic beads, which were subsequently delivered to a detection chamber housing multiple sensors. The assay was designed to accurately detect CD62L in diluted serum with a limit of detection (LOD) of 0.25 ng mL-1 and a dynamic range of 0.25-100 ng mL-1. The microfluidic array gave significantly better accuracy and higher sensitivity than a standard ELISA kit, which was shown to be subject to significant systematic error at high and low concentration ranges. 31 serum samples from patients with varying grades of bladder cancer and cancer-free controls were analyzed by the immunoarray and ELISA, and the CD62L levels correlated. This work establishes a new accurate assay for determining CD62L levels and highlights the potential of this protein as a biomarker for detecting locoregional progression of bladder cancer.


Asunto(s)
Biomarcadores de Tumor/sangre , Selectina L/sangre , Neoplasias de la Vejiga Urinaria/sangre , Animales , Bovinos , Técnicas Electroquímicas/métodos , Ensayo de Inmunoadsorción Enzimática/métodos , Humanos , Límite de Detección , Técnicas Analíticas Microfluídicas/métodos
2.
Biosens Bioelectron ; 77: 188-93, 2016 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-26406460

RESUMEN

Herein we report a low cost, sensitive, supercapacitor-powered electrochemiluminescent (ECL) protein immunoarray fabricated by an inexpensive 3-dimensional (3D) printer. The immunosensor detects three cancer biomarker proteins in serum within 35 min. The 3D-printed device employs hand screen printed carbon sensors with gravity flow for sample/reagent delivery and washing. Prostate cancer biomarker proteins, prostate specific antigen (PSA), prostate specific membrane antigen (PSMA) and platelet factor-4 (PF-4) in serum were captured on the antibody-coated carbon sensors followed by delivery of detection-antibody-coated Ru(bpy)3(2+) (RuBPY)-doped silica nanoparticles in a sandwich immunoassay. ECL light was initiated from RuBPY in the silica nanoparticles by electrochemical oxidation with tripropylamine (TPrA) co-reactant using supercapacitor power and ECL was captured with a CCD camera. The supercapacitor was rapidly photo-recharged between assays using an inexpensive solar cell. Detection limits were 300-500f gmL(-1) for the 3 proteins in undiluted calf serum. Assays of 6 prostate cancer patient serum samples gave good correlation with conventional single protein ELISAs. This technology could provide sensitive onsite cancer diagnostic tests in resource-limited settings with the need for only moderate-level training.


Asunto(s)
Conductometría/instrumentación , Suministros de Energía Eléctrica , Inmunoensayo/instrumentación , Mediciones Luminiscentes/instrumentación , Impresión Tridimensional , Análisis por Matrices de Proteínas/instrumentación , Capacidad Eléctrica , Diseño de Equipo , Análisis de Falla de Equipo , Miniaturización , Reproducibilidad de los Resultados , Sensibilidad y Especificidad
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