New positive control peptide pools: CEFRAS Global
Published: January 21, 2025
Updated: May 27, 2025
7 minute read
Authored by: Tyler Sandberg
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 the PepPool: CEFRAS Global family of peptide pools.
These pools set a whole new standard for performance and coverage in ELISpot and FluoroSpot experiments. Consisting of carefully selected T cell epitopes derived from 18-22 different pathogens, the CEFRAS Global pools are designed for effective T cell stimulation across a wide range of HLA types and ethnic backgrounds. Antigen-specific positive controls are crucial in your T cell assays to ensure that your T cells are functionally active and can respond to peptide stimulation. Pair these pools with a mitogen as well and you're good to go!
The CEFRAS Global peptide pool family consists of three different peptide pools:
- PepPool: CEFRAS Global (CD8), human
- PepPool: CEFRAS Global (CD4), human
- PepPool: CEFRAS Global (CD4+CD8), human
All of the epitopes selected for these peptide pools were carefully chosen after studying published literature that identified highly immunogenic regions of common pathogen antigens. With our expertise in immunology and immunoassays, we understand the art of identifying and designing quality peptide pools with epitopes that should elicit a high number of T cells in your samples, enriching the robustness of your assay and data.
In the following sections, we’ll go into the details of each of these pools with performance data and benchmarking against other popular positive control pools. Can I use this peptide pool in flow cytometry? Yes, you can! While we are still generating more flow cytometry validation data, we know that this peptide pool should also work in intracellular cytokine staining assays. Test it out in ICS and let us know what you think!
PepPool: CEFRAS Global (CD8), human
When you need a positive control peptide pool to elicit robust CD8 T cell responses, look no further! Consisting of 210 carefully selected MHC class-I restricted T cell epitopes derived from 18 different pathogens, the CEFRAS Global (CD8) pool has been designed for effective T cell stimulation across a broader range of HLA types and ethnic backgrounds.
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.
Performance data
We designed this pool with quality in mind and we don’t launch a pool until we are 100% satisfied with its functionality and ensure it works with real samples. Our benchmarking studies highlight the significant edge this CD8 peptide pool holds over other peptide pool suppliers as well. When tested against Company A’s popular positive control peptide pool, our new pool consistently achieved stronger T cell responses in both high and medium-low responders. For example, in the IFN-γ ELISpot assays, the CEFRAS Global (CD8) 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.
PepPool: CEFRAS Global (CD4), human
Up next is the CEFRAS Global (CD4) peptide pool. Designed to stimulate superior CD4 T cell responses in a broad range of HLA types and ethnic backgrounds, this pool is made up of 85 MHC class-II restricted peptides covering 20 pathogens, including SARS-CoV-2, RSV, MTB, and Haemophilus influenzae, not covered by other major CD4 peptide pools.
PepPool CEFRAS global (CD4), human consists of antigenic epitopes from several pathogens that we are commonly exposed to or vaccinated against. Our T cell analyses have shown that this pool delivers higher responses in a greater number of donors compared to other popular CD4 control pools. As immunologists, we're picky when it comes to selecting immunodominant peptides, which is why we carefully studied the literature to help influence the design of the CEFRAS pools.
Performance data
As mentioned before, we designed this pool with quality in mind and made sure it works in actual assays with actual samples before launching. The data below shows the results from 250,000 PBMCs per well from 18 donors stimulated with either CEFRAS Global (CD4), our CEFTA pool, or Company A's MHC-II restricted peptide pool and incubated overnight. IFN-γ ELISpot results revealed a significant increase in SFU from PBMCs stimulated with either CEFRAS Global (CD4) or CEFTA pools compared to Company A's popular CD4 peptide pool, indicating superior performance.
And we didn't stop our analyses there. We used the same stimulation setup as before, but with IFN-γ/IL-4/IL-5 FluoroSpot as the readout. We found that CEFRAS Global (CD4) outperformed Competitor A's MHC-II restricted peptide pool in both IFN-γ and IL-4 SFU. This increased pathogen coverage and specific selection of immunodominant epitopes make our new pool a dependable choice in your important experiments, driving your research forward.
PepPool: CEFRAS Global (CD4+CD8), human
If you're looking for a "one-size-fits-all" peptide-pool for stimulating both CD8 and CD4 T cells as positive controls, then CEFRAS Global (CD4+CD8), human is the solution for you! Although similar with some overlap, this pool is not simply the previously described CD4 and CD8 CEFRAS Global pools combined, but a different pool designed specifically for the stimulation of both CD4 and CD8 T cells. CEFRAS Global (CD4+CD8) consists of 203 peptides covering 22 pathogens, including RSV, MTB, and Yellow Fever Virus, which are not often found in other CD4+CD8 pools.
Performance data
Just like our individual CD4 and CD8 CEFRAS Global pools, the CD4+CD8 combination pool has been meticulously designed to stimulate both T cell families in a large range of donors. IFN-γ ELISpot was used to compare T cell responses in healthy donors after stimulation with either CEFRAS Global (CD4+CD8) or Company A's CD4+CD8 positive control pool. The results revealed a significant increase in SFU from PBMCs stimulated with Mabtech's CEFRAS Global (CD4+CD8) compared to Company A's popular CD4+CD8 peptide pool.
We continued our evaluation of our new CD4+CD8 peptide pool using both FluoroSpot Plus: Human IFN‑γ/IL‑2/TNF‑α and FluoroSpot Plus: Human IFN‑γ/IL‑4/IL‑5 kits. We found that all donors showed an increase in secretion of the tested cytokines, demonstrating proper function of the CEFRAS Global (CD4+CD8) in experimental conditions.
Our new family of positive control peptide pools, CEFRAS Global, is designed to deliver reliable stimulation across a broad range of HLA types and samples from diverse ethnic backgrounds. 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 Global pools simplify the workflow for immunoassay experiments. With high-quality capping 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, these pools are the ideal solutions.
Ready to try them out yourself? Contact your account manager or click below!