This document summarizes research on using a filtration assisted luminometric ELISA (FAL-ELISA) to detect foot-and-mouth disease virus (FMDV) non-structural proteins in formulated vaccines as an alternative to animal testing. The researchers were able to extract the FMDV 3B protein from the aqueous phase of spiked FMDV-oil vaccines using FAL-ELISA. This demonstrates the feasibility of detecting FMDV non-structural proteins in formulated vaccines with FAL-ELISA. The researchers conclude that FAL-ELISA is a promising candidate to replace animal testing for quality control of FMD vaccines, which could be useful for vaccine banks and regulatory bodies.

1. 1EuFMD | Open Session special edition | #OS20se
CECILIA TURCO
Team: Florencia Mansilla, María Cruz Miraglia and Alejandra Capozzo.
INTA and CONICET
Institute of Virology and Technical Innovations. Buenos Aires
ARGENTINA
FILTRATION ASSISTED LUMINOMETRIC ELISA
(FAL-ELISA) APPLIED TO THE DETECTION OF
FOOT-AND-MOUTH DISEASE VIRUS NON-
STRUCTURAL PROTEINS IN FORMULATED
VACCINES

2. 2
FAL-ELISA: principle
MD | Open Session special edition | #OS20se
YY
Y Y YY Y Y
MONOCLONAL
ANTIBODY
anti-3B
SAMPLES (vaccine
extraction) and
CONTROLS
HRP Mab anti-
3ABC
Y Y Y
YY
LUMINOL/
PEROXIDE
SAMPLES DETECTION
20 min15 min 40 min
Y Y Y
AIM: assess the feasibility of detecting FMDV-3B protein in formulated vaccines using the
IPC FMDV kit

3. 3
EXTRACTION PROTOCOL
PROOF OF PRINCIPLE: detection of
NSP in formulated vaccines
• Extraction of the aqueous
phase from the different
vaccine preparations with
chloroform (1:1 V/V).
• Performed by triplicates
during 2, 4 or 24h to
evaluate 3ABC stability at
4°C
3ABC-spiked
vaccine
3ABC in the aqueous phase extracted from spiked
FMDV-oil vaccines using FAL-ELISA
Plate validation
C30/NIL 4611,92 >20
C10/NIL 1519,05 >5
C30/C10 3,04 >1,5
Quantitation
Sample RLU Result
Spiked 50 ng 364030 POS
Spiked 50 ng (1:2) 90620 POS
Spiked 10 ng 54780 POS
NIL 37480 NEG

4. 4
✓ FAL-ELISA could be used for the detection of FMDV non-structural proteins in
formulated vaccines.
✓ FAL-ELISA is a promising candidate for replacing the use of animals for the
control of FMD-vaccine purity, that can be particularly useful for vaccine
banks and regulatory bodies.
Conclusion

5. eufmdvirtual.com
8, 10, 15, 17 December 2020
EuFMD / Open Session