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

Journal of Immunological Methods
Janine D MullerLin-Fa Wang

Abstract

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 (a...Continue Reading

References

Nov 1, 1989·Analytical Biochemistry·S C Gill, P H von Hippel
Jun 1, 1996·Immunotechnology : an International Journal of Immunological Engineering·R J KerschbaumerG Himmler
Nov 11, 1999·Journal of Molecular Recognition : JMR·D G Myszka
Jul 6, 2004·Journal of Virological Methods·Jody D BerryFrank Plummer
Oct 10, 2006·Analytical Biochemistry·Giuseppe A PapaliaDavid G Myszka

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Citations

Dec 3, 2014·Biologicals : Journal of the International Association of Biological Standardization·Gaurav K SharmaBramhadev Pattnaik

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Methods Mentioned

BETA
ELISA
PCR
phage display
biosensor
size exclusion chromatography
surface plasmon resonance
chip
size
Gel Filtration
C-ELISA

Software Mentioned

Scrubber

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