Production and characterization of single-chain antibody (scFv) against 3ABC non-structural protein in Escherichia coli for sero-diagnosis of Foot and Mouth Disease virus
| Autor: | Bramhadev Pattnaik, Rakesh Matura, Sonalika Mahajan, Saravanan Subramaniam, Jajati K. Mohapatra, Gaurav Sharma |
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| Rok vydání: | 2014 |
| Předmět: |
Buffaloes
Population Enzyme-Linked Immunosorbent Assay Bioengineering Antibodies Viral Sensitivity and Specificity Applied Microbiology and Biotechnology Serology Immunoenzyme Techniques Mice Osmotic Pressure Escherichia coli medicine Animals Serologic Tests Cloning Molecular education Pharmacology education.field_of_study Hybridomas General Immunology and Microbiology Foot-and-mouth disease biology Antibodies Monoclonal Reproducibility of Results Viral Vaccines General Medicine medicine.disease biology.organism_classification Virology Recombinant Proteins ROC Curve Foot-and-Mouth Disease Virus Polyclonal antibodies Foot-and-Mouth Disease Monoclonal biology.protein Cattle Electrophoresis Polyacrylamide Gel Foot-and-mouth disease virus Antibody Single-Chain Antibodies Biotechnology |
| Zdroj: | Biologicals. 42:339-345 |
| ISSN: | 1045-1056 |
| Popis: | Differentiation of Foot-and-Mouth Disease infected from vaccinated animals is essential for effective implementation of vaccination based control programme. Detection of antibodies against 3ABC non-structural protein of FMD virus by immunodiagnostic assays provides reliable indication of FMD infection. Sero-monitoring of FMD in the large country like India is a big task where thousands of serum samples are annually screened. Currently, monoclonal or polyclonal antibodies are widely used in these immunodiagnostic assays. Considering the large population of livestock in the country, an economical and replenishable alternative of these antibodies was required. In this study, specific short chain variable fragment (scFv) antibody against 3B region of 3ABC poly-protein was developed. High level of scFv expression in Escherichia coli system was obtained by careful optimization in four different strains. Two formats of enzyme immunoassays (sandwich and competitive ELISAs) were optimized using scFv with objective to differentiate FMD infected among the vaccinated population. The assays were statistically validated by testing 2150 serum samples. Diagnostic sensitivity/specificity of sandwich and competitive ELISAs were determined by ROC method as 92.2%/95.5% and 89.5%/93.5%, respectively. This study demonstrated that scFv is a suitable alternate for immunodiagnosis of FMD on large scale. |
| Databáze: | OpenAIRE |
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