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.
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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