Development of a Novel, Ultra-rapid Biosensor for the Qualitative Detection of Hepatitis B Virus-associated Antigens and Anti-HBV, Based on 'Membrane-engineered' Fibroblast Cells with Virus-Specific Antibodies and Antigens
| Autor: | Antonis Perdikaris, Nikos Alexandropoulos, Spiridon Kintzios |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
HBsAg
Bioelectric Recognition Assay Cell biosensor Membrane-engineering Hepatitis viruses Vero cells lcsh:Chemical technology medicine.disease_cause Biochemistry Article Virus Analytical Chemistry Blood serum Antigen medicine lcsh:TP1-1185 Electrical and Electronic Engineering Instrumentation Hepatitis B virus biology virus diseases Virology Molecular biology digestive system diseases Atomic and Molecular Physics and Optics biology.protein Vero cell Antibody Biosensor |
| Zdroj: | Sensors; Volume 9; Issue 3; Pages: 2176-2186 Sensors (Basel, Switzerland) Sensors, Vol 9, Iss 3, Pp 2176-2186 (2009) |
| ISSN: | 1424-8220 |
| DOI: | 10.3390/s90302176 |
| Popis: | A novel miniature cell biosensor detection system for the detection of Hepatis B virus (HBV)-associated antigens and anti-HBV is described. The biosensor is based on "membrane-engineered" Vero fibroblast cells immobilized in an alginate matrix. The membrane-engineering process involved the electroinsertion of anti-HBV specific antibodies (anti-HBs, anti-HBe) or antigens (HBsAg) in the membranes of the Vero cells. The attachment of a homologous antigen to the electroinserted antibody (or, respectively, of the antibody to the electroinserted antigen) triggered specific changes to the cell membrane potential that were measured by appropriate microelectrodes, according to the principle of the Bioelectric Recognition Assay (BERA). The sensor was used for screening 133 clinical blood serum samples according to a double-blind protocol. Considerably higher sensor responses were observed against HBV-positive samples, compared with responses against negative samples or samples positive for heterologous hepatitis viruses such as Hepatitis C (HCV) virus. Detection of anti-HBs antibodies was made possible by using a biosensor based on immobilized Vero cells bearing the respective antigen (HBsAg). The observed response was rapid (45 sec) and quite reproducible. Fluorescence microscopy observations showed that attachment of HBV particles to cells membrane-engineered with anti-HBs was associated with a decrease of [Ca(2+)]cyt. The perspectives for using the novel biosensor as a qualitative, rapid screening, high throughput assay for HBV antigens and anti-HBs in clinical samples is discussed. |
| Databáze: | OpenAIRE |
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