New positive control peptide pool: CEFRAS Global (CD8), human
Published: January 21, 2025
Updated: January 23, 2025
Proper positive controls are the key to success in any immunoassay. Peptide pools combining several common antigens that should elicit T cell responses in the general population have served as positive controls for years. Recently, we’ve seen that popular peptide pool controls work great in certain populations but not at all in others. That’s why we set out to create a new positive control peptide pool that would work in a larger range of populations with diverse ethnic backgrounds. Introducing PepPool: CEFRAS Global (CD8), human.
This pool sets a whole new standard for performance and coverage in ELISpot, FluoroSpot, and flow cytometry experiments. Consisting of 210 carefully selected MHC class-I restricted T cell epitopes derived from 18 different pathogens, the CEFRAS Global (CD8) pool ensures effective T cell stimulation across a wide range of HLA types and ethnic backgrounds.
| PepPool: CEFRAS (CD8), human design | |||
|---|---|---|---|
| Pathogen | No. peptides | Pathogen | No. peptides |
| SARS-CoV-2 | 36 | Vaccinia virus | 7 |
| Human herpesvirus 4 (EBV) | 30 | Yellow fever | 6 |
| Influenza A | 25 | Toxoplasma gondii | 4 |
| Human herpesvirus 5 (CMV) | 24 | Human herpesvirus 2 (HSV-2) | 3 |
| Human adenovirus (Ad5) | 18 | Human herpesvirus 6 (HHV6) | 3 |
| Human herpesvirus 1 (HSV-1) | 18 | Human papilloma virus (HPV 16 L1) | 3 |
| Human herpesvirus (VZV) | 10 | JC polyomavirus | 2 |
| Respiratory syncytial virus (RSV) | 10 | Coxackievirus B4 | 1 |
| Mycobacterium tuberculosis | 9 | Measles virus | 1 |
The PepPool CEFRAS global (CD8), human consists of antigenic epitopes from a number of pathogens that we are commonly exposed to or vaccinated against. Our analyses have shown that this pool delivers higher responses in a greater number of donors compared to other popular control pools. This superior performance stems from our meticulous selection process, which ensures a comprehensive representation of antigens and pathogens. The pool consists of epitopes from Coxsackievirus B4, Human adenovirus 5, Human herpesvirus 1-6, Human papillomavirus, Influenza A, JC polyomavirus, Measles virus, Mycobacterium tuberculosis, Respiratory syncytial virus, SARS-CoV-2, Toxoplasma gondii, Vaccinia virus, and Yellow fever virus. The pool not only covers a wide spectrum of infectious agents but also enables researchers to generate consistent and reliable results, even when working with diverse donor samples.
We designed this pool with quality in mind. We don’t launch a pool until we are 100% satisfied with its quality and ensure it works with real samples. Take this ICS experiment for example. We stimulated human PBMCs with the CEFRAS pool and measured the resulting production of IFN-γ and TNF-α. The results were consistent across the board showing significant increases in production of all three of these analytes after stimulation with the new pool.
Human PBMCs were stimulated with the CEFRAS (CD8) peptide pool (2 μg/ml) for 12 hours in the presence of Brefeldin A. Cells were stained for viability using ViaDye Red, followed by surface marker staining: CD14, CD19, CD56, CD3, CD4, CD8, CCR7, CD45RA, and CX3CR1. After fixation and permeabilization with 4% paraformaldehyde and saponin, intracellular cytokines (IFN-ү, TNF-ɑ, IL-2) were stained. Flow cytometry was performed on a Cytek Northern Lights (Violet, Blue, Red lasers).
Our benchmarking studies highlight the significant edge this peptide pool holds over competitors as well. When tested against Company A’s positive control peptide pool, our product consistently achieved stronger T cell responses in both high and medium-low responders. For example, in the IFN-γ ELISpot assays, our pool produced substantially higher spot-forming units (SFUs) across all donor categories. Similarly, in FluoroSpot assays, the frequency of cytokine-secreting cells stimulated by our pool outperformed competing products. This higher sensitivity and broader applicability make our pool a dependable choice for researchers striving for accurate, reproducible results.
Whether you're working with samples from Europe, Asia, Africa, or the Americas, our peptide pool delivers reliable stimulation. This global reach empowers researchers to confidently design experiments that reflect real-world scenarios, bridging the gap between preclinical research and clinical applications. In addition to its technical benefits, the CEFRAS (CD8) pool simplifies the workflow for immunoassay experiments. With high-quality synthesis and rigorous validation, the pool reduces variability and saves valuable time in assay setup. For researchers seeking the highest level of confidence in their data, this pool is the ideal solution.
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