recombinant hace 2  (ACROBiosystems)

Journal: Clinical Immunology (Orlando, Fla.)

Article Title: Seroprevalence and SARS-CoV-2 cross-reactivity of endemic coronavirus OC43 and 229E antibodies in Finnish children and adults

doi: 10.1016/j.clim.2021.108782

Figure Lengend Snippet: SARS-CoV-2 neutralization potency of the pre-pandemic hCoV antibodies. Individual sera ( n = 10) positive (grey columns) or negative (white columns) for SARS-CoV-2 receptor-binding domain (RBD) and/or spike-specific antibodies were used to block spike (a) or RBD (b) proteins binding to immobilized human angiotensin-converting enzyme 2 (ACE-2) in a surrogate neutralization assay. RBD-specific monoclonal antibody (mAb) was used as positive blocking control (black columns). The columns represent the binding inhibition percent (%) for each sample was calculated as 100% - (optical density [OD] of ACE-2 binding in the presence of serum/OD of ACE-2 maximum binding × 100%). The numerical values above/inside columns illustrate the OD-values obtained from spike (a) or RBD (b) -specific IgG ELISAs.

Article Snippet: Ninety-six well plates (Corning) were coated overnight with human ACE-2 (hACE-2, aa18–740) Fc chimera recombinant protein (1 μg/ml in PBS, Invitrogen) and the plates were blocked with 5% milk in PBS for one hour at RT.

Techniques: Neutralization, Binding Assay, Blocking Assay, Inhibition