Name: PepTivator® SARS-CoV-2 Prot_S
Availability: InStock

Product data and images for PepTivator
^®
SARS-CoV-2 Prot_S

Figures

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 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

**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

**Recovered COVID-19 patients show elevated levels of IFN-γ–producing CD3**⁺ **T cells upon stimulation with SARS‑CoV‑2 PepTivator Peptide Pools.**
Samples of a healthy donor and a recovered COVID-19 patient were stimulated for 4 h with the indicated SARS-CoV-2 PepTivator Peptide Pools (Prot_N, Prot_M, Prot_S, or a mix of all three) in the presence of BFA. As negative control, samples were left untreated without (w/o) antigen. Subsequently, T cell lineage surface markers and intracellular cytokines were stained. The presented plots are exemplary data showing IFN-γ
⁺
CD3
⁺
T cells.

Figure 3

Recovered COVID-19 patients show elevated levels of IFN-γ–producing CD3⁺ T cells upon stimulation with SARS‑CoV‑2 PepTivator Peptide Pools.

Samples of a healthy donor and a recovered COVID-19 patient were stimulated for 4 h with the indicated SARS-CoV-2 PepTivator Peptide Pools (Prot_N, Prot_M, Prot_S, or a mix of all three) in the presence of BFA. As negative control, samples were left untreated without (w/o) antigen. Subsequently, T cell lineage surface markers and intracellular cytokines were stained. The presented plots are exemplary data showing IFN-γ

⁺

CD3

⁺

T cells.

Figure 4

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 5

**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 5

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

Stimulate T cells reactive to the spike protein of the SARS-CoV-2 Wuhan wild-type using PepTivator Peptide Pools. PepTivators are pools of lyophilized peptides, consisting mainly of 15-mer sequences with 11 amino acids overlap. PepTivator SARS-CoV-2 Prot_S is a pool of lyophilized peptides, covering the immunodominant sequence domains of the surface (or spike) glycoprotein (“S”) of SARS-Coronavirus 2 (GenBank MN908947.3, Protein QHD43416.1). The PepTivator SARS-CoV-2 Prot_S contains the sequence domains aa 304-338, 421-475, 492-519, 683-707, 741-770, 785-802, and 885 – 1273 (sequence end).

In contrast, PepTivator SARS-CoV-2 Prot_S Complete covers all functional domains (aa 5–1273), Prot_S1 the complete N-terminal S1 domain (aa 1–692) 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.

SARS-CoV-2 Prot_S stands for the surface glycoprotein of SARS-CoV-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 initiate, which enables the viral genome to enter the host cell. 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 selected immunodominant sequence domains of the spike protein (aa 304–338, 421–475, 492–519, 683–707, 741–770, 785–802, and 885–1273).

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:

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).

More information regarding the work with antigen-specific T cells can be found in the T cell application section.

Resources for PepTivator
^®
SARS-CoV-2 Prot_S

Documents and Protocols

Scientific posters

Detection of Humoral and Cellular Responses to SARS-CoV-2 in COVID-19 Convalescents

App notes/customer reports

Rapid antigen-reactive T cell enrichment (Rapid ARTE) - Workflow protocol

App notes/customer reports

Fast and efficient detection of SARS-CoV-2–reactive T cells via antigen-specific stimulation

References for PepTivator
^®
SARS-CoV-2 Prot_S

Publications

Sekine, T. et al. (2020) Robust T Cell Immunity in Convalescent Individuals with Asymptomatic or Mild COVID-19. Cell 183(1): 158-168
Au, L. et al. (2021) Cytokine release syndrome in a patient with colorectal cancer after vaccination with BNT162b2. Nat Med 27(8): 1362-1366
Shroff, R. T. et al. (2021) Immune Responses to COVID-19 mRNA Vaccines in Patients with Solid Tumors on Active, Immunosuppressive Cancer Therapy. Nat Med 27(11): 2002-2011
Stephenson, E. et al. (2021) Single-cell multi-omics analysis of the immune response in COVID-19. Nat Med 27(5): 904-916
Sir Karakus, G. et al. (2021) Preclinical efficacy and safety analysis of gamma-irradiated inactivated SARS-CoV-2 vaccine candidates. Sci Rep 11(1): 5804
Cooper, R. S. et al. (2021) Rapid GMP-Compliant Expansion of SARS-CoV-2-Specific T Cells From Convalescent Donors for Use as an Allogeneic Cell Therapy for COVID-19. Front Immunol 11: 598402
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 12(doi: 10.3389/fimmu.2021.708820)
Cotugno, N. et al. (2021) Virological and immunological features of SARS-CoV-2-infected children who develop neutralizing antibodies. Cell Rep 34(11): 108852
Riou, C. et al. (2020) Rapid, simplified whole blood-based multiparameter assay to quantify and phenotype SARS-CoV-2 specific T cells. Eur. Respir. J. 59(1): 2100
Chan, Y.-H. et al. (2021) Asymptomatic COVID-19: disease tolerance with efficient anti-viral immunity against SARS-CoV-2. EMBO Mol. Med. 13(6)
Fischer, D. S. et al. (2021) Single-cell RNA sequencing reveals ex vivo signatures of SARS-CoV-2-reactive T cells through ‘reverse phenotyping’. Nat Commun. 26(12): 4515
Grau-Expósito, J. et al. (2021) Peripheral and lung resident T cell responses against SARS-CoV-2. Nat Commun. 12(1): 3010
Jung, J. H. et al. (2021) SARS-CoV-2-specific T Cell Memory is Sustained in COVID-19 Convalescents for 8 Months with Successful Development of Stem Cell-like Memory T Cells. Nat Commun. 12(1): 4043
Kramer, K. J. et al. (2022) Single-Cell Profiling of the Antigen-Specific Response to BNT162b2 SARS-CoV-2 RNA Vaccine. Nat Commun. 13(1): 3466
Kusnadi, A. et al. (2021)
Severely ill COVID-19 patients display augmented functional properties in SARS-CoV-2-reactive CD8
⁺
T cells.
Sci Immunol 6: eabe4782
Anft, M. et al. (2020) COVID-19 progression is potentially driven by T cell immunopathogenesis. medRxiv : doi: 10.1101/2020.04.28.20083089
Leung, W. et al. (2020) Successful manufacturing of clinical-grade SARS-CoV-2–specific T cells for adoptive cell therapy. medRxiv : doi: 10.1101/2020.04.24.20077487
Willingham, S. B. et al. (2020) Characterization and Phase 1 Trial of a B Cell Activating Anti-CD73 Antibody for the Immunotherapy of COVID-19. medRxiv : doi: 10.1101/2020.09.10.20191486
Li, Z. et al. (2020) SARS-CoV-2-specific T cell memory is long-lasting in the majority of convalsecent COVID-19 individuals. bioRxiv : doi: 10.1101/2020.11.15.383463
Nanishi, E. et al. (2021) Alum:CpG adjuvant enables SARS-CoV-2 RBD-induced protection in aged mice and synergistic activation of human elder type 1 immunity. bioRxiv : doi: 10.1101/2021.05.20.444848
Singh, D. K. et al. (2020) SARS-CoV-2 infection leads to acute infection with dynamic cellular and inflammatory flux in the lung that varies across nonhuman primate species. bioRxiv : doi: 10.1101/2020.06.05.136481
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
Silva-Cayetano, A. et al. (2021) A booster dose enhances immunogenicity of the COVID-19 vaccine candidate ChAdOx1 nCoV-19 in aged mice. Med (N Y) 2(3): 243-262
Demaret, J. et al. (2020) Severe SARS‐CoV‐2 patients develop a higher specific T‐cell response. Clin Transl Immunology 9(12): e1217
Ferreras, C. et al. (2020) SARS-CoV-2-Specific Memory T Lymphocytes From COVID-19 Convalescent Donors: Identification, Biobanking, and Large-Scale Production for Adoptive Cell Therapy. Front Cell Dev Biol. 9: 9:620730
Anft, M. et al. (2021) Detection of pre-existing SARS-CoV-2-reactive T cells in unexposed renal transplant patients. J Nephrol. 34(4): 1025-1037
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
Fendler, A. et al. (2021) Adaptive immunity to SARS-CoV-2 in cancer patients: The CAPTURE study. Nat Cancer 2: 1321-1337
Mazzoni, A. et al. (2021) First dose mRNA vaccination is sufficient to reactivate immunological memory to SARS-CoV-2 in ex COVID-19 subjects. J. Clin. Invest. 131(12): doi: 10.1101/2021.03.05.21252590
Riou, C. et al. (2021) Profile of SARS-CoV-2-specific CD4 T cell response: Relationship with disease severity and impact of HIV-1 and active Mycobacterium tuberculosis co-infection. J. Clin. Invest. 131(12): doi: 10.1172/JCI149125
Sattler, A. et al. (2020) SARS-CoV-2-specific T cell responses and correlations with COVID-19 patient predisposition. J. Clin. Invest. 130(12): 6477-6489
Bonifacius, A. et al. (2021) COVID-19 immune signatures reveal stable antiviral T cell function despite declining humoral responses. Immunity 54(2): 340-354
Shomuradova, A. S. et al. (2020) SARS-CoV-2 epitopes are recognized by a public and diverse repertoire of human T cell receptors. Immunity 53(6): 1245-1257
Habel, J. R. et al. (2020)
Suboptimal SARS-CoV-2-specific CD8
⁺
T cell response associated with the prominent HLA-A*02:01 phenotype.
Proc. Natl. Acad. Sci. U.S.A. 117(3): 24384-24391
Harrington, P. et al. (2021) Single dose of BNT162b2 mRNA vaccine against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) induces neutralising antibody and polyfunctional T-cell responses in patients with chronic myeloid leukaemia. Br J Haematol 194(6): 999-1006
Meckiff, J. et al. (2020)
Imbalance of Regulatory and Cytotoxic SARS-CoV-2-Reactive CD4
⁺
T Cells in COVID-19.
Cell 183(5): 1340-1353

Data and images