Detection of inducible nitric oxide synthase using a suite of electrochemical, fluorescence, and surface plasmon resonance biosensors
| Autor: | Omowunmi A. Sadik, Naumih M. Noah, Saamia Alam |
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| Rok vydání: | 2010 |
| Předmět: |
Silver
Biophysics Analytical chemistry Nitric Oxide Synthase Type II Enzyme-Linked Immunosorbent Assay Biochemistry Blood Substitutes Limit of Detection medicine Animals Humans Surface plasmon resonance Fluorescent Antibody Technique Indirect Molecular Biology Detection limit Chromatography biology Chemistry Cell Biology Electrochemical Techniques Surface Plasmon Resonance Human serum albumin Fluorescence Primary and secondary antibodies Nitric oxide synthase Polyclonal antibodies Calibration biology.protein Biosensor Oxidation-Reduction medicine.drug |
| Zdroj: | Analytical biochemistry. 413(2) |
| ISSN: | 1096-0309 |
| Popis: | A suite of biosensors for rapid detection of inducible nitric oxide synthase (iNOS) is described. First, a metal-enhanced electrochemical detection (MED) sensor, which relied on the redox properties of a silver monolayer, was developed. The linear detection range was between 8.64 × 10 −2 and 5.4 × 10 1 ng/ml with a detection limit of 1.69 × 10 −4 ng/ml. This method was compared with surface plasmon resonance (SPR) biosensors in which polyclonal mouse anti-iNOS was covalently immobilized onto a gold surface using an iNOS antigen. The linear detection range recorded was between 3.37 × 10 1 and 5.4 × 10 −2 ng/ml with a detection limit of 2 × 10 −3 ng/ml. Finally, an ultrasensitive portable capillary (UPAC) fluorescence immunosensor, in which a mouse anti-iNOS antibody was covalently immobilized onto the inner surface of a capillary and a rabbit anti-iNOS antibody was employed as the secondary antibody, was developed. The resulting signals were found to be directly proportional to iNOS concentrations between 1.52 × 10 −1 and 1.52 × 10 −2 ng/ml with a detection limit of 1.05 × 10 −3 ng/ml. These immunosensors exhibit low cross-reactivity toward potential interferents such as human serum albumin and ovalbumin. The SPR and UPAC biosensors were validated using simulated blood spiked with recombinant iNOS, resulting in recoveries of 85% and 88.5%, respectively. The research presented in this article could potentially provide new ways of detecting NO for diagnostic and biomarker purposes in medical research. |
| Databáze: | OpenAIRE |
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