Improvement of a recombinant antibody-based serological assay for foot-and-mouth disease virus

Autor: Michelle Wilkins, Janine D. Muller, Hans G. Heine, Olan Dolezal, Meng Yu, Lin-Fa Wang, Adam J. Foord
Rok vydání: 2010
Předmět:
IPTG
isopropyl-beta-D-thiogalactoside
Cost-Benefit Analysis
Recombinant Fusion Proteins
viruses
Epitope mapping
Immunology
C-ELISA
competition enzyme-linked immunosorbent assay
Enzyme-Linked Immunosorbent Assay
DIVA test
Viral Nonstructural Proteins
Antibodies
Viral
Protein Engineering
Binding
Competitive
Article
GST
glutathione S-transferase
scFv
Virus
Epitope
FMDV
PCR
polymerase chain reaction
SARS
Severe Acute Respiratory Syndrome
Alkaline phosphatase
DIVA
differentiation of infected from vaccinated animals
Animals
Immunology and Allergy
Serologic Tests
CRAb
chicken recombinant antibody
PAGE
polyacrylamide gel electrophoresis
Aphthovirus
biology
Reproducibility of Results
Viral Vaccines
scFv
single chain variable fragment
biology.organism_classification
Virology
Fusion protein
Diva
AP
alkaline phosphatase
Foot-and-Mouth Disease Virus
Foot-and-Mouth Disease
IBDV
infectious bursal disease virus
biology.protein
FMDV
foot-and-mouth disease virus
Binding Sites
Antibody
Foot-and-mouth disease virus
Antibody
Zdroj: Journal of Immunological Methods
ISSN: 0022-1759
DOI: 10.1016/j.jim.2009.11.004
Popis: Differentiating foot-and-mouth disease virus (FMDV) antibodies generated during a natural infection from those due to vaccination (DIVA) is crucial for proving freedom from disease after an outbreak and allowing resumption of trade in livestock products. The World Organisation for Animal Health (OIE) recommends that FMDV vaccines are composed of inactivated virus that has been purified to remove non-structural viral proteins. Such purified vaccines primarily induce antibodies to viral structural proteins, whereas replicating virus stimulates host antibodies specific for both structural and non-structural proteins. The current preferred FMDV DIVA test is a competitive ELISA (C-ELISA) designed to detect antibodies to the non-structural protein 3ABC. Previously, we described the development of an FMDV DIVA test based entirely on recombinant proteins (the recombinant detecting antibody and the 3ABC coating antigen) produced in Escherichia coli. In this study, we have determined the precise binding site of the recombinant detecting antibody to a conserved sequence within the 3B region of the 3ABC protein, replaced the original E-tag of the detecting antibody with two in-house tags and engineered a direct antibody-reporting enzyme (alkaline phosphatase) fusion protein. These modifications have further improved the DIVA test, providing great potential for large scale production and uptake due to its simplicity, reproducibility and low cost.
Databáze: OpenAIRE
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