Human Anti-Staphylococcus aureus IsdB Recombinant Antibody (clone STAU-281)

Biolayer interferometry was used to detect whether the mAbs blocked binding of hemoglobin to NEAT2. Biotinylated hemoglobin was loaded onto streptavidin-coated biosensor tips before association with either IsdB alone, IsdB mixed with mAb, mAb alone, or kinetic buffer. Error bars represent standard deviations. P values were determined by an unpaired t test. This experiment was performed at least 3 independent times.

Bennett, M. R., Dong, J., Bombardi, R. G., Soto, C., Parrington, H. M., Nargi, R. S., ... & Crowe Jr, J. E. (2019). Human VH1-69 gene-encoded human monoclonal antibodies against Staphylococcus aureus IsdB use at least three distinct modes of binding to inhibit bacterial growth and pathogenesis. MBio, 10(5), e02473-19.

(d to f) S. aureus heme-dependent in vitro growth curves were performed in 96-well plates over 32 h. S. aureus strain Newman was subcultured at a 1:200 dilution into RPMI medium with ethylenediamine-N,N'-bis(2-hydroxyphenylacetic acid) and normalized to an OD600 of 1.0.

Bennett, M. R., Dong, J., Bombardi, R. G., Soto, C., Parrington, H. M., Nargi, R. S., ... & Crowe Jr, J. E. (2019). Human VH1-69 gene-encoded human monoclonal antibodies against Staphylococcus aureus IsdB use at least three distinct modes of binding to inhibit bacterial growth and pathogenesis. MBio, 10(5), e02473-19.

Seven-week-old female BALB/c mice were inoculated retro-orbitally with a suspension of S. aureus strain Newman at an OD600 of 0.4. Mice were given IGHV1-69-encoded full-length wild-type IgG LALA Fc variant (L234A/L235A) IgG antibodies by the intraperitoneal route. The hearts, livers, and kidneys of the infected mice were harvested after 96 h. Statistical significance was evaluated by analysis of variance (ANOVA) using multiple comparisons. All other comparisons, including comparison of LALA mAbs to the isotype control, were not significant.

Bennett, M. R., Dong, J., Bombardi, R. G., Soto, C., Parrington, H. M., Nargi, R. S., ... & Crowe Jr, J. E. (2019). Human VH1-69 gene-encoded human monoclonal antibodies against Staphylococcus aureus IsdB use at least three distinct modes of binding to inhibit bacterial growth and pathogenesis. MBio, 10(5), e02473-19.