Polyclonal goat antimouse igg hrp

Purified recombinant protein and isolated LPS were separated on a one-dimensional SDS-PAGE and electroblotted onto PVDF membranes. FtHU and LPS were detected using a polyclononal rabbit anti FTS_0886 serum (Moravian Biotechnology) and a mouse monoclonal anti-LPS FB11 antibody (Abcam, AB2033), respectively. As secondary antibodies the polyclonal swine antirabbit IgG/HRP (Dako, P0399) and polyclonal goat antimouse IgG/HRP (Dako, P0447) were used. Chemiluminescence detection was employed using a BM Chemiluminescence Blotting Substrate (POD) while following the manufacturer's instructions (Roche, 11500694001). LPS staining was done using Pro-Q Emerald 300 Gel Stain kit (Invitrogen, P20495). Images of gels were collected using a UV transilluminator (Vilber Lourmat, Eberhardzell, Germany).

The neutralization potencies of nirsevimab, MEDI8897*, MEDI8897*-TM, and palivizumab were evaluated against recombinant RSV A and RSV B viruses in a microneutralization susceptibility assay. Each mAb was serially diluted in 96-well plates prior to the addition of a fixed concentration of RSV A000 or RSV B000 at a median tissue culture infectious dose (TCID₅₀) of 500 per well and was incubated for 1 hour followed by the addition of HEp-2 cells. After 5 days of incubation at 37°C, RSV-infected cells were fixed and stained with anti-RSV mAb (Millipore, Cat# MAB8262 Clone 133-1H at 1:5000 dilution) and a horseradish peroxidase-labelled detection antibody (Dako polyclonal goat anti-mouse IgG-HRP Cat# P0447 at 1:4000 dilution). Tetramethylbenzidine substrate was added and the OD_450nm was measured. Half-maximal inhibitory concentrations (IC₅₀) were calculated using 4PL nonlinear curve fitting on the GraphPad Prism (version 9.4.0) software suite and compared to the IC₅₀ values of reference viruses on the same plate, to determine fold-change in IC₅₀ values.