PepTivator
®
SARS-CoV-2 Prot_S1 is a pool of lyophilized peptides, consisting mainly of 15-mer sequences with 11 amino acids overlap, covering the N-terminal S1 domain of the surface glycoprotein ("S") of SARS-Coronavirus 2 (GenBank MN908947.3, Protein QHD43416.1). The PepTivator SARS-CoV-2 Prot_S1 contains the aa sequence 1–692 of the surface glycoprotein, 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)

Data and images for
PepTivator
®
SARS-CoV-2 Prot_S1

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_S1

Overview

PepTivator
®
SARS-CoV-2 Prot_S1 is a pool of lyophilized peptides, consisting mainly of 15-mer sequences with 11 amino acids overlap, covering the N-terminal S1 domain of the surface glycoprotein ("S") of SARS-Coronavirus 2 (GenBank MN908947.3, Protein QHD43416.1). The PepTivator SARS-CoV-2 Prot_S1 contains the aa sequence 1–692 of the surface glycoprotein, 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_S+ parts of the C-terminal S2 domain (aa 689–895). 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.
The surface glycoprotein of SARS CoV-2, the "spike 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. The spike protein consists of two functional domains: The S1 domain contains the surface binding site to the ACE2 receptor and the S2 subunit mediates membrane fusion. Cleavage at the S1/S2 boundary (aa-residues 685 and 686) via cellular proteases is crucial to prime the protein for the entry to the host cell.¹ The cleavage site distinguishes SARS-CoV-2 from SARS-CoV spike protein sequence, and is thought to be an important feature not only for its zoonotic potential
²
but also for increased SARS-CoV-2 transmissibility
¹,⁴
. Thus, the spike proteins, and especially the S1 domain, 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_S1 covers the N-terminal S1 domain of the spike protein (aa 1–692). 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_S+ parts of the C-terminal S2 domain (aa 689–895). 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.

Applications

PepTivator
®
SARS-CoV-2 Prot_S1 – 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:
  • 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.
  • 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).

References for
PepTivator
®
SARS-CoV-2 Prot_S1

Publications

  1. Haun, B. K. et al. (2020) CoVaccine HT™ Adjuvant Potentiates Robust Immune Responses to Recombinant SARS-CoV-2 Spike S1 Immunization Front Immunol 11(doi: 10.3389)
  2. 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)
  3. Chan, Y-H. et al. Asymptomatic COVID-19: disease tolerance with efficient anti-viral immunity against SARS-CoV-2 EMBO Mol. Med. (doi: 10.15252/emmm.202114045)
  4. Anft, M. et al. (2020) COVID-19 progression is potentially driven by T cell immunopathogenesis medRxiv - the preprint server for health sciences (doi.org/10.1101/2020.04.28.20083089 )
  5. 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)
  6. Thieme, C. J. et al. (2020) Robust T Cell Response Toward Spike, Membrane, and Nucleocapsid SARS-CoV-2 Proteins Is Not Associated with Recovery in Critical COVID-19 Patients. Cell Rep Med. 1(6): 100092
  7. 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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