Table 1.
Description of the control and vaccinated groups.
Fig 1.
The depiction of the experimental design and use of an Influenza A hemagglutinin (HA) monovalent vectored vaccine in the porcine animal model vaccinated then boosted 28 days later.
HA-specific IgG and IgA antibodies were evaluated in serum, milk and colostrum (Figure created with BioRender.com. Byrne, J. (2025) https://biorender.com/g36e786).
Fig 2.
The project design of the antibody isolation protocol leading to the evaluation of HA-specific antibody response post-vaccination via in-house HA ELISA.
A. Selecting a resin dependent on desired antibody isotype to isolate. B. Pig-derived sample collection to be diluted, plated, and incubated with the resin to bind the antibody. C. Centrifugation of the stacked plates with the elution buffer and collection of the isolated antibodies in the bottom collection plate. D. The eluted products are moved to individual, clean filter tubes for concentration and changing the storage buffer at the same time. E. The eluted antibody is moved and stored in a clean collection microcentrifuge tube. F. Each isolated antibody sample is pipetted onto the nanodrop spectrophotometer to record the published concentration in mg/mL. G. The isolated antibody is diluted to a specified concentration and plated in a precoated low-bind ELISA plate for optical density (OD) analysis, confirming vaccine-induced antibody specificity. (Figure created with BioRender.com. Byrne, J. (2025) https://biorender.com/s11c541).
Fig 3.
Isolation of swine immunoglobulins from serum, colostrum, and milk significantly reduced background OD450 values.
A. ELISA results of vaccinated and control groups comparing whole serum, 2 weeks post-boost (1:100 dilution), to antibodies (50 µg/mL) isolated from serum using Protein G for IgG and CaptureSelect for IgA B. ELISA results of vaccinated and control groups comparing whole milk, 2 weeks post-boost (1:100 dilution), to antibodies (50 µg/mL) isolated from milk using Protein G for IgG and CaptureSelect for IgA C. ELISA results of vaccinated and control groups comparing whole colostrum, 2 weeks post-prime (1:100 dilution), to antibodies (50 µg/mL) isolated from colostrum using Protein G for IgG and CaptureSelect for IgA. OD 450nm. Mean ± SD. N = 4. Statistics: Unpaired t test. * p < 0.05 **p < 0.01 ***p < 0.001 **** p < 0.0001.