This application protocol describes how SARS-CoV-2–specific antibodies can be detected in plasma or serum samples of COVID-19 convalescents using Miltenyi Biotec's biotinylated recombinant SARS-CoV-2 proteins in a bead-based flow cytometric assay. The humoral response of SARS-CoV-2−convalescent patients is an essential background information to analyze cellular responses, such as antigen-specific B cells, more in detail.
Recombinant proteins
Table 1: Exemplary calculation of the concentrations of reconstituted recombinant proteins
Recombinant protein variant | Concentration after reconstitution (mg/mL) | MW (kDa) | Corresponding molar concentration (µM) |
Recombinant SARS-CoV-2 RBD (HEK)-Biotin | 0.1 | 33.8 | 2.96 |
Recombinant SARS-CoV-2 RBD (Insect cells)-Biotin | 0.1 | 34.2 | 2.92 |
Recombinant SARS-CoV-2 Nucleoprotein-Biotin | 0.1 | 49.1 | 2.04 |
Recombinant SARS-CoV-2 Spike-Trimer (HEK)-Biotin | 0.1 | 418.0 | 0.24 |
Recombinant SARS-CoV-2 Spike-S2 (insect cells) | 0.1 | 61.2 | 1.63 |
Recombinant SARS-CoV-2 Spike-Prot (HEK)- Biotin | 0.1 | 136.0 | 0.74 |
Recombinant Human ACE2 (HEK)-Biotin | 0.1 | 87.2 | 1.15 |
Recombinant Human ACE2 (insect cells) | 0.1 | 72.8 | 1.37 |
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