Gene/Protein Disease Symptom Drug Enzyme Compound
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We examined Automated Chemiluminescence System ACS: 180 for CEA assay, especially about confidence of the functional sensitivity. Serum CEA levels of normal subjects and patients were measured not only with ACS: 180 but also with immunoradiometric assay (CEA kit [Daiichi] II). Using ACS: 180 assay for CEA, intra-assay coefficients of variation (C.V.s) were 4.06% at 1.97 ng/ml, 1.62% at 17.3 ng/ml, and 2.28% at 81.3 ng/ml. Interassay C.V.s were 4.02% at 1.99 ng/ml, 1.21% at 16.5 ng/ml, and 3.44% at 79.4 ng/ml. The detection limit (functional sensitivity) of ACS: 180 assay for CEA was 0.4 ng/ml by the precision profile. The coefficient of correlation of 164 sera, which CEA values were among 0.4-100 ng/ml (working range of ACS: 180), between with ACS: 180 and with CEA kit [Daiichi] II was R = 0.942, y = 1.07 x +0.09 (p < 0.05). Seven (13.2%) normal subjects (n = 53) and 45 (34.1%) patients with malignant diseases (n = 132) had higher serum CEA levels than the cut off value (2.5 ng/ml). The sera of 82 malignant patients who had both pre- and post-operation sample were determined. Sixty-five patients of them were decrease after operation. These results of ACS: 180 resembled to that of CEA kit [Daiichi] II. We conclude that ACS: 180 for CEA assay was precise enough to measure below the cut off value, and had good performance of its speed (short incubation) and convenience.
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PMID:[Immunochemiluminometric assay (Automated Chemiluminescence System ACS: 180) for CEA]. 852 39

This study demonstrates a facile and feasible strategy toward the development of advanced electrochemical immunosensors based on chemically functionalized magnetic mesoporous organic-inorganic hybrid nanomaterials, and the preparation, characterization, and measurement of relevant properties of the immunosensor for detection of carcinoembryonic antigen (CEA, as a model analyte) in clinical immunoassays. The as-prepared nanomaterials composed of a magnetic mesoporous NiCo(2)O(4) nanosheet, an interlayer of Nafion/thionine organic molecules and a nanogold layer show good adsorption properties for the attachment of horseradish peroxidase-labeled secondary anti-CEA antibody (HRP-anti-CEA). With a sandwich-type immunoassay format, the functional bionanomaterials present good analytical properties to facilitate and modulate the way it was integrated onto the electrochemical immunosensors, and allows the detection of CEA at a concentration as low as 0.5 pg/mL. Significantly, the immunosensor could be easily regenerated by only using an external magnet without the need of any dissociated reagents. Importantly, the as-synthesized magnetic mesoporous NiCo(2)O(4) nanomaterials could be further extended for detection of other biomarkers or biocompounds.
ACS Appl Mater Interfaces 2011 Apr
PMID:Magnetic mesoporous organic-inorganic NiCo2O4 hybrid nanomaterials for electrochemical immunosensors. 2144 71

A novel and in situ amplified immunoassay strategy with quadruple signal amplification was designed for highly efficient electrochemical detection of low-abundance proteins (carcinoembryonic antigen, CEA, as a model) by using nanogold-functionalized DNAzyme concatamers with redox-active intercalators. To construct such an in situ amplification system, streptavidin-labeled gold nanoparticles (AuNP-SA) were initially used for the labelling of initiator strands (S0) and detection antibody (mAb2) with a large ratio (mAb2-AuNP-S0), and then two auxiliary DNA strands S1 and S2 were designed for in situ propagation of DNAzyme concatamers with the hemin/G-quadruplex format. The quadruple signal amplification was implemented by using the avidin-biotin chemistry, nanogold labels, DNA concatamers, and DNAzymes. In the presence of target CEA, the sandwiched immunocomplex was formed between the immobilized primary antibodies on the electrode and the conjugated detection antibodies on the mAb2-AuNP-S0. The carried S0 initiator strands could progress a chain reaction of hybridization events between alternating S1/S2 DNA strands to form a nicked double-helix. Upon addition of hemin, the hemin-binding aptamers could be bound to form the hemin/G-quadruplex-based DNAzymes. The formed double-helix DNA polymers could cause the intercalation of numerous electroactive methylene blue molecules. During the electrochemical measurement, the formed DNAzymes could catalyze the reduction of H2O2 in the solution to amplify the electrochemical signal of the intercalated methylene blue. Under optimal conditions, the electrochemical immunoassay exhibited a wide dynamic range of 1.0 fg mL(-1) to 20 ng mL(-1) toward CEA standards with a low detection limit of 0.5 fg mL(-1). Intra-assay and inter-assay coefficients of variation (CV) were less than 8.5% and 11.5%, respectively. No significant differences at the 0.05 significance level were encountered in the analysis of 14 clinical serum specimens between the developed immunoassay and commercialized electrochemiluminescent (ECL) method for detection of CEA.
ACS Appl Mater Interfaces 2013 Apr 10
PMID:Nanogold-functionalized DNAzyme concatamers with redox-active intercalators for quadruple signal amplification of electrochemical immunoassay. 2348 56

Point-of-care (POC) testing has the potential to enable rapid, low-cost, and large-scale screening. POC detection of a multiplexed biomarker panel can facilitate the early diagnosis of non-small cell lung cancer (NSCLC) and, thus, may allow for more timely surgical intervention for life-saving treatment. Herein, we report the nanoporous glass (NPG) integrated volumetric bar-chart chip (V-Chip) for POC detection of the three NSCLC biomarkers CEA, CYFRA 21-1, and SCCA, by the naked eye. The 3D nanostructures in the NPG membrane efficiently increase the number of binding sites for antibodies and decrease the diffusion distance between antibody and antigen, enabling the low detection limit and rapid analysis time of the NPG-V-Chip. We utilized the NPG-V-Chip to test the NSCLC biomarker panel and found that the limit of detection can reach 50 pg/mL (10-fold improvement over the original V-Chip), and the total assay time can be decreased from 4 to 0.5 h. We then detected CEA in 21 serum samples from patients with common cancers, and the on-chip results showed good correlation with the clinical results. We further assayed 10 lung cancer samples using the device and confirmed the results obtained using conventional ELISA methods. In summary, the NPG-V-Chip platform has the ability of multiplex, low detection limit, low cost, lack of need for accessory equipment, and rapid analysis time, which may render the V-Chip a useful platform for quantitative POC detection in resource-limited settings and personalized diagnostics.
ACS Nano 2016 Jan 26
PMID:Nanoporous Glass Integrated in Volumetric Bar-Chart Chip for Point-of-Care Diagnostics of Non-Small Cell Lung Cancer. 2669 Jul 45

This article describes a photochemical approach for independently patterning multiple proteins to an inert substrate, particularly for studies of cell adhesion. A photoactivatable chloropyrimidine ligand was employed for covalent immobilization of SnapTag fusion proteins on self-assembled monolayers of alkanethiolates on gold. A two-step procedure was used: first, patterned UV illumination of the surface activated protein capture ligands, and second, incubation with a SnapTag fusion protein bound to the surface in illuminated regions. Two different fluorescent proteins were patterned in registry with features of 400 nm in size over a 1 mm2 area. An example is given wherein an anti-carcinoembryonic antigen (anti-CEA) scFv antibody was patterned to direct the selective attachment of a human cancer cell line that express the CEA antigen. This method enables the preparation of surfaces with control over the density and activity of independently patterned proteins.
ACS Appl Mater Interfaces 2018 Nov 28
PMID:Photoactivatable Reaction for Covalent Nanoscale Patterning of Multiple Proteins. 3037 16

Coordination polymers (CPs) as fascinating materials have been explored in a number of fields due to their diverse properties. In this work, we demonstrate the feasibility of CPs in the facile fabrication of multifunctional composites for establishing an immunoassay. To this end, a zinc(II)-based CP (ZnCP) with adenine as a bridge ligand was employed to integrate with alkaline phosphatase (ALP) and anticarcinoembryonic antigen (anti-CEA) antibody, which produces ALP/anti-CEA@ZnCPs. Benefiting from the adaptive inclusion property of ZnCPs, the integrated ALP and anti-CEA can maintain their original catalytic activity and capture ability to target antigen, respectively. This allows the ALP/anti-CEA@ZnCPs to be a detection antibody for performing an immunoassay. Meanwhile, ZnCP as a host can effectively protect the loaded ALP and anti-CEA against harsh environments. On this basis, by using iron(II)-phenanthroline complex as a signal amplifier, a colorimetric immunoassay for CEA detection was developed, and a low detection limit of 21.1 pg/mL has been achieved. This immunoassay was successfully applied to determine CEA levels in serum samples with good recovery and precision. We believe that this study can not only provide a new method for CEA detection but also open up a new way for the rational design and fabrication of multifunctional composites.
ACS Appl Mater Interfaces 2019 Nov 20
PMID:A Colorimetric Immunoassay Based on Coordination Polymer Composite for the Detection of Carcinoembryonic Antigen. 3167 5