1.
A novel sandwich-type photoelectrochemical immunosensor based on Ru(bpy)32+ and Ce-CdS co-sensitized hierarchical ZnO matrix and dual-inhibited polystyrene@CuS-Ab2 composites.
Fan, D, Liu, X, Bao, C, Feng, J, Wang, H, Ma, H, Wu, D, Wei, Q
Biosensors & bioelectronics. 2019;:124-131
Abstract
A novel and sensitive sandwich-type photoelectrochemical (PEC) immunosensor was developed for the quantitative detection of β-amyloid protein (Aβ). A ITO electrode was sequentially coated with hierarchical porous zinc oxide (ZnO) microspheres with a large specific area, sensitized with tris(bipyridine)ruthenium(II) ion (Ru(bpy)32+) to achieve high visible light absorption, and modified with cerium-doped cadmium sulfide (Ce-CdS) nanoparticles to enhance the PEC response. Under the stimulation of visible light and ascorbic acid as an efficient electron donor, the photoelectric signal of ZnO/Ru(bpy)32+/Ce-CdS was 70 times that of pure ZnO. The amino-functionalized polystyrene (PS) microspheres coated with copper sulfide (CuS) was linked with a secondary antibody (Ab2) for the first time for the Aβ detection by the immunosensor. The good insulation and steric resistance of the as-prepared polystyrene@CuS-Ab2 (PS@CuS-Ab2) composite significantly weakened the photocurrent response of the immunosensor in the specific immune recognition. Under the optimal conditions, the quantitative detection of Aβ was achieved within the range of 0.001-100 ng/mL with the detection limit of 0.37 pg/mL. In addition, the PEC immunosensor is easy to make, stable and selective, which has provided a good experimental platform for the detection of disease biomarkers.
2.
Engineering regulable Escherichia coli beta-galactosidases as biosensors for anti-HIV antibody detection in human sera.
Ferrer-Miralles, N, Feliu, JX, Vandevuer, S, Müller, A, Cabrera-Crespo, J, Ortmans, I, Hoffmann, F, Cazorla, D, Rinas, U, Prévost, M, et al
The Journal of biological chemistry. 2001;(43):40087-95
Abstract
The activity of engineered, peptide-displaying enzymes is modulated by binding to specific anti-peptide antibodies. This new concept of a quantitative antibody detection system allows test kits to be set up for fast diagnosis of infectious diseases. To develop a quick and homogeneous assay for the detection of human immunodeficiency virus (HIV) infection, we have explored two acceptor sites of the bacterial Escherichia coli beta-galactosidase for the accommodation of HIV antigenic peptides. Two overlapping epitopes (namely P1 and P2) from the gp41 envelope glycoprotein, contained in different sized peptides, were inserted in the vicinity of the enzyme active site to generate a set of hybrid, enzymatically active beta-galactosidases. Regulable enzymes of different responsiveness to monoclonal antibody binding were generated with both acceptor sites tested. These biosensors were also sensitive to immune sera from HIV-infected patients. Modeling data provide insight into the structural modifications in the vicinity of the active site induced by peptide insertion that strongly affect the responsiveness of the engineered proteins through different parameters of their catalytic properties.