Marv angola gpdtm

Assays for antibody effector functions were adapted from previously established protocols.²⁰ (link) Post–challenge NHP sera were inactivated by gamma-irradiation (4 MRad)¹⁸ (link) and removed from the maximum containment laboratory according to RML SOPs approved by the RML IBC. Recombinant MARV-Angola GPdTM (IBT Bioservices) was tethered to Fluospheres NutrAvidin-Microspheres yellow-green or red (Thermo Fisher Scientific, Waltham, MA) using the EZ-link Micro Sulfo-NHS-LC-Biotinylation kit (Thermo Fisher Scientific).

Post–challenge NHP sera were inactivated by gamma-irradiation (4 MRad)¹⁸ (link) and removed from the maximum containment laboratory according to SOPs approved by the RML IBC. The MARV GP-specific IgM titers in serum samples were determined at 1:250 dilution using ELISA kits following manufacturer's instructions (Alpha Diagnostics, San Antonio, TX). The MARV GP and VP40 IgG ELISAs were developed in-house. MARV-Angola GPdTM was obtained from IBT Bioservices (Gaithersburg, MD) and MARV VP40 was purified from transfected 293T cell supernatant. Nunc Maxisorp Immuno plates (Thermo Fisher Scientific) were coated with 50 μL of 1 μg/mL of antigen in PBS o/n and ELISAs were performed as described previously.¹⁹ (link) The optical density (OD) at 405 nm was measured using a GloMax® explorer (Promega). The OD values were normalized to the baseline samples obtained with naïve NHP serum and the cutoff value was set as the mean OD plus three times the standard deviation of the blank.