20-S2S-TCg-L

SARS-CoV-2 recombinant Spike (SCoV2-rS) glycoprotein trimeric ectodomain from mammalian CHO cells, lyophilized

Description: SARS-CoV-2 recombinant Spike (SCoV2-rS) glycoprotein ectodomain expressed as secreted trimeric protein in mammalian (hamster) CHO cells. UniProtKB sequence accession number: P0DTC2 (SPIKE_SARS2); expressed ectodomain includes amino acids (aa) 1-1208. Whole expression construct includes: full-length SCoV2-S ectodomain (aa 1 – 1208) w/o transmembrane and cytoplasmic aa, furin cleavage site “RRAR” mutated to “GSAS”, C-terminal GCN4 trimerization motif fused to protein sequence, then follows thrombin cleavage site, Strep-tag II and His6-tag. Two mutations (K986P and V987P) were introduced into SCoV2-S sequence to stabilize the trimer in the pre-fusion conformation. SCoV2-rS glycoprotein was purified from culture medium under native (non-denaturing) conditions using convenient chromatographic techniques.

Form: Lyophilized in PBS (pH 7.4).

Storage: Shipped at ambient temperature (shipment costs to Europe: 70 €). Lyophilized products may be stored at 2-8°C or at -20°C for longer periods.

Reconstitution: add 100 μl of deionized water for reconstitution of 100 µg of SCoV2-rS in each separate vial. Check lyophilized material to be completely dissolved before use. Reconstituted protein is completely soluble (remained in solution after centrifugation at 10000 x g for 20 min.).

Concentration of reconstituted product: 1.0 mg/ml in PBS.

Purity: >90 % as determined by SDS-PAGE.

Image: SDS-PAGE showing SCoV2-rS product 20-S2S-TCg-L at approximately 160 kDa (1, 3 and 5 μg/lane).

Applications (tested): ELISA, Western blot, SDS-PAGE.

Quality: According to the expressed construct, purified SCoV2-rS protein should be trimer in the stabilized prefusion conformation. Full-length trimeric protein enables detection of more antibodies against SCoV2-S, than shorter protein constructs (e.g., RBD or S1). This product 20-S2S-TCg-L showed similar results in ELISA with COVID-19 positive and negative patient sera, as analogous protein construct formulated in glycerol stock (see product 20-S2S-TCg-G information), but a lesser number of individual sera was tested.

Reactivity of SCoV2-rS glycoprotein (current batch of product 20-S2S-TCg-L) with specific IgG in ELISA using pools of Covid-19 positive and negative sera:

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