PepTivator
®
SARS-CoV-2 Prot_S+ is a pool of lyophilized peptides, consisting mainly of 15-mer sequences with 11 amino acids (aa) overlap, covering the sequence domain aa 689–895 of the surface glycoprotein ("S") of SARS-Coronavirus 2 (GenBank MN908947.3, Protein QHD43416.1), also termed "spike" protein.
In contrast, PepTivator SARS-CoV-2 Prot_S Complete covers all functional domains (aa 5–1273), Prot_S covers selected immunodominant sequence domains of the spike protein (aa 304–338, 421–475, 492–519, 683–707, 741–770, 785–802, and 885–1273) and Prot_S1 the complete N-terminal S1 domain (aa 1–692). The complete S2 domain (and parts of the S1 domain:

Data and images for
PepTivator
®
SARS-CoV-2 Prot_S+

Figures

Figure 1

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Schematic alignment of the spike glycoprotein and different SARS‑CoV‑2 PepTivator Peptide Pools based on this protein.
PepTivator SARS‑CoV‑2 Prot_S covers the predicted immunodominant domains of the SARS‑CoV‑2 spike glycoprotein (protein S), PepTivator SARS‑CoV‑2 Prot_S1 covers the N-terminal S1 domain, PepTivator SARS‑CoV‑2 Prot_S+ covers a part of the C-terminal S2 domain, and PepTivator SARS-CoV-2 Prot_S Complete covers the whole protein sequence of the spike protein without the first 4 amino acids of the signal peptide.

Figure 1

Schematic alignment of the spike glycoprotein and different SARS‑CoV‑2 PepTivator Peptide Pools based on this protein.
PepTivator SARS‑CoV‑2 Prot_S covers the predicted immunodominant domains of the SARS‑CoV‑2 spike glycoprotein (protein S), PepTivator SARS‑CoV‑2 Prot_S1 covers the N-terminal S1 domain, PepTivator SARS‑CoV‑2 Prot_S+ covers a part of the C-terminal S2 domain, and PepTivator SARS-CoV-2 Prot_S Complete covers the whole protein sequence of the spike protein without the first 4 amino acids of the signal peptide.

Figure 2

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Exemplary analysis of SARS-CoV-2–specific CD4+ and CD8+ T cells.
A) PBMCs were stimulated with a mix of PepTivator SARS-CoV-2 Prot_N, Prot_M, and Prot_S or unstimulated as negative control. The data shows CD154 and TNF-α for CD4
+
T cells and TNF-α and IFN-γ for CD8
+
T cells. B/C). Either PepTivators covering the complete sequence of the nucleoprotein (N; PepTivator SARS-CoV-2 Prot_N), the membrane protein
(M; PepTivator SARS-CoV-2 Prot_M), the spike protein (S; PepTivator SARS-CoV-2 Prot_S, Prot_S1, and Prot_S+), or the mix of all were used to stimulate SARS-CoV-2–reactive T cells. Frequencies of CD154+ within CD4
+
T cells and TNF-α
+
within CD8
+
T cells are shown. In the quantitative analysis (B) each dot corresponds to one donor and in the heat maps (C) SARS-CoV-2–reactive T cell frequencies measured for each donor are color coded.

Figure 2

Exemplary analysis of SARS-CoV-2–specific CD4+ and CD8+ T cells.
A) PBMCs were stimulated with a mix of PepTivator SARS-CoV-2 Prot_N, Prot_M, and Prot_S or unstimulated as negative control. The data shows CD154 and TNF-α for CD4
+
T cells and TNF-α and IFN-γ for CD8
+
T cells. B/C). Either PepTivators covering the complete sequence of the nucleoprotein (N; PepTivator SARS-CoV-2 Prot_N), the membrane protein
(M; PepTivator SARS-CoV-2 Prot_M), the spike protein (S; PepTivator SARS-CoV-2 Prot_S, Prot_S1, and Prot_S+), or the mix of all were used to stimulate SARS-CoV-2–reactive T cells. Frequencies of CD154+ within CD4
+
T cells and TNF-α
+
within CD8
+
T cells are shown. In the quantitative analysis (B) each dot corresponds to one donor and in the heat maps (C) SARS-CoV-2–reactive T cell frequencies measured for each donor are color coded.

Figure 3

View details
Quantitative analysis of spike protein–specific CD4+ T cells in convalescent COVID-19 donors.
Quantitative analysis of SARS‑CoV‑2–specific CD4
+
T cells of convalescent COVID-19 donors highlights that similar numbers of activated T cells can be observed whether the mix of three PepTivator Peptide Pools
(PepTivator SARS-CoV-2 Prot_S, PepTivator SARS-CoV-2 Prot_S1, and PepTivator SARS-CoV-2 Prot_S+) or the single PepTivator SARS-CoV-2 Prot_S Complete is used.

Figure 3

Quantitative analysis of spike protein–specific CD4+ T cells in convalescent COVID-19 donors.
Quantitative analysis of SARS‑CoV‑2–specific CD4
+
T cells of convalescent COVID-19 donors highlights that similar numbers of activated T cells can be observed whether the mix of three PepTivator Peptide Pools
(PepTivator SARS-CoV-2 Prot_S, PepTivator SARS-CoV-2 Prot_S1, and PepTivator SARS-CoV-2 Prot_S+) or the single PepTivator SARS-CoV-2 Prot_S Complete is used.

Figure 4

View details
Stimulation with SARS-CoV-2 PepTivator Peptide Pools reveals the presence of virus-specific CD4+ T cells after vaccination.
A whole blood sample of a vaccinated donor was stimulated with the mix of three PepTivator Peptide Pools (PepTivator SARS-CoV-2 Prot_S, PepTivator SARS-CoV-2 Prot_S1, and PepTivator SARS-CoV-2 Prot_S+) or with PepTivator SARS-CoV-2 Prot_S Complete alone.
Both, the applied mix of peptide pools as well as SARS-CoV-2 Prot_S Complete cover the complete sequence of the spike protein. As negative control, the sample was left unstimulated. T cell lineage surface markers and intracellular cytokines were stained, and cells were analyzed using a MACSQuant Analyzer 16. The presented plots are exemplary data showing CD154 and TNF-α for pregated CD4
+
T cells.

Figure 4

Stimulation with SARS-CoV-2 PepTivator Peptide Pools reveals the presence of virus-specific CD4+ T cells after vaccination.
A whole blood sample of a vaccinated donor was stimulated with the mix of three PepTivator Peptide Pools (PepTivator SARS-CoV-2 Prot_S, PepTivator SARS-CoV-2 Prot_S1, and PepTivator SARS-CoV-2 Prot_S+) or with PepTivator SARS-CoV-2 Prot_S Complete alone.
Both, the applied mix of peptide pools as well as SARS-CoV-2 Prot_S Complete cover the complete sequence of the spike protein. As negative control, the sample was left unstimulated. T cell lineage surface markers and intracellular cytokines were stained, and cells were analyzed using a MACSQuant Analyzer 16. The presented plots are exemplary data showing CD154 and TNF-α for pregated CD4
+
T cells.

Specifications for
PepTivator
®
SARS-CoV-2 Prot_S+

Overview

PepTivator
®
SARS-CoV-2 Prot_S+ is a pool of lyophilized peptides, consisting mainly of 15-mer sequences with 11 amino acids (aa) overlap, covering the sequence domain aa 689–895 of the surface glycoprotein ("S") of SARS-Coronavirus 2 (GenBank MN908947.3, Protein QHD43416.1), also termed "spike" protein.
In contrast, PepTivator SARS-CoV-2 Prot_S Complete covers all functional domains (aa 5–1273), Prot_S covers selected immunodominant sequence domains of the spike protein (aa 304–338, 421–475, 492–519, 683–707, 741–770, 785–802, and 885–1273) and Prot_S1 the complete N-terminal S1 domain (aa 1–692). The complete S2 domain (and parts of the S1 domain: aa 304–338, 421–475, and 492–519) is covered, when PepTivator SARS-CoV-2 Prot_S and Prot_S+ are combined.
In vitro
stimulation of antigen-specific T cells with PepTivator Peptide Pools causes the secretion of effector cytokines and the up-regulation of activation markers, which then allow the detection and isolation of antigen-specific T cells.

Detailed product information

Background information

SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2), also known as novel coronavirus 2019-nCoV, causes fever, severe respiratory illness, and can lead to life threatening pneumonia. The first cases of this disease, termed COVID-19 for coronavirus disease 2019, have been detected in December 2019 in Wuhan, China. SARS-CoV-2 Prot_S stands for the surface glycoprotein of SARSCoV-2, the “spike protein”. This protein is responsible for the recognition and binding of the coronavirus to the host cell. Once SARS-CoV-2 has bound to the ACE2 receptor of the host cell, fusion of viral envelope and host cell membrane starts, which enables the viral genome to enter the host cell
1,2
. Thus, the spike proteins are crucial for the infection of cells with coronaviruses and have been suggested as possible target for vaccine development.
The PepTivator
®
SARS-CoV-2 Prot_S+ covers the gaps in the sequence mapping between aa 689 and 895 of the PepTivator SARS-CoV-2 Prot_S. Both PepTivators can be combined to cover the complete C-terminal S2 domain of the spike protein, which is cleaved from the S1-domain by a furin protease
3,4
. In contrast, PepTivator SARS-CoV-2 Prot_S Complete covers all functional domains (aa 5–1273) and Prot_S1 the complete N-terminal S1 domain (aa 1–692).

Applications

PepTivator
®
SARS-CoV-2 Prot_S+ – research grade has been specifically developed for
in vitro
stimulation of SARS-CoV-2–specific T cells. Peptides of 15 amino acids in length and 11 amino acids overlap represent an optimized solution for stimulating both CD4
+
and CD8
+
T cells in various applications, including:
  • The PepTivator SARS-CoV-2 Prot_S+ can be combined with PepTivator SARS-CoV-2 Prot_S (# 130-126-700; # 130-126-701) for covering the complete C-terminal S2-domain of the spike protein (and parts of the S1 domain: aa 304–338, 421–475, and 492–519).
  • Detection and analysis of SARS-CoV-2–specific CD4+ and CD8+ effector/memory T cells in PBMCs by MACS® Cytokine Secretion Assays, intracellular cytokine staining, or other technologies.
  • Isolation of viable SARS-CoV-2–specific CD4+ T cells with the CD154 MicroBead Kit, or of viable CD4+ and CD8+ T cells using MACS Cytokine Secretion Assay – Cell Enrichment and Detection Kits. Subsequently, cells can be expanded for generation of T cell lines.
  • Generation of SARS-CoV-2–specific CD4+ and CD8+ effector / memory T cells from naive T cell populations.
  • Pulsing of antigen-presenting cells, e.g. for research on dendritic cell vaccination.

References for
PepTivator
®
SARS-CoV-2 Prot_S+

Publications

  1. Strafella, C. et al. (2021) Case Report: Sars-CoV-2 Infection in a Vaccinated Individual: Evaluation of the Immunological Profile and Virus Transmission Risk Front Immunol (doi: 10.3389/fimmu.2021.708820)
  2. Mazzoni, A. et al. (2021) First dose mRNA vaccination is sufficient to reactivate immunological memory to SARS-CoV-2 in ex COVID-19 subjects medRxiv - the preprint server for health sciences (doi.org/10.1101/2021.03.05.21252590)
  3. Aiello, A. et al. (2021) Spike is the most recognized antigen in the whole-blood platform in both acute and convalescent COVID-19 patients Int J Infect Dis 106: 338-347

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®
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