Precision Medicine

Cellular Phenotyping Services

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Cellular Phenotyping in Immune Monitoring and Beyond: Exploring Diverse Applications with PeploBio

Cellular phenotyping, a cornerstone technique in immune monitoring, offers versatile applications that encompass not only traditional immune system analysis but also specialised assays such as CAR-T cell analysis and stem cell phenotyping.

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PeploBio's Expertise in Cellular Phenotyping

At PeploBio, we specialise in providing comprehensive cellular phenotyping solutions tailored to our clients' specific research needs. Our team of experienced scientists utilises state-of-the-art technologies, including the BD FACSLyric clinical flow cytometer, to offer a wide range of services. With the BD FACSuite™ Application, we ensure assay standardisation and compliance with regulatory requirements such as 21 CFR Part 11 and electronic record integrity, providing our clients with confidence in the quality of their research data (1) .

Our services encompass

Immune Cell Characterisation

Accurate identification and quantification of immune cell subsets, including T cells, Bcells, NK cells, dendritic cells, and monocytes/macrophages.

Stem Cell Analysis

Precise enumeration and characterisation of stem cell populations using a panel of markers tailored for stem cell identification and functional assessment.

CAR-T Cell Analysis

Evaluation of CAR-T cell properties, including CAR expression, activation status, and functional capabilities, to assess treatment efficacy and patient response.

Cellular phenotyping serves as a powerful tool for dissecting the complexities of the immune system and beyond. With its broad range of applications and advanced technologies, PeploBio remains committed to providing cutting-edge cellular phenotyping solutions to advance scientific research and improve human health.Speak to one of our scientists today.

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The Evolution of  
Cellular Phenotyping Techniques

Over the years, technological advancements have revolutionised the field of cellular phenotyping, enabling researchers to delve deeper into the complexity of cellular dynamics (3). Flow cytometry, one of the cornerstone techniques in cellular phenotyping, has undergone significant advancements, allowing for high-throughput analysis of multiple parameters simultaneously. Modern flow cytometers, such as the BD FACSLyric used at PeploBio, offer enhanced sensitivity, resolution, and automation, making them indispensable tools for immune monitoring and cell-based research (4) .

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Marker-Based Identification of Cell Populations

Central to cellular phenotyping is the identification and characterisation of specific cell populations using a combination of surface markers and functional assays.For example, T cells can be distinguished based on the expression of CD3, CD4, and CD8 markers (5), while B cells are characterised by the presence of CD19 and CD20 (6) . Natural killer (NK) cells, on the other hand, can be identified by the absence of CD3 and the expression of CD56 (7).

In addition to immune cell subsets, cellular phenotyping also enables the identification of stem cell populations and CAR-T cells using specific marker panels. Stem cells, with their unique regenerative properties, express markers such as CD34, CD45, and CD117 (8). These markers facilitate the precise enumeration and characterisation of stem cell populations, providing insights into their differentiation potential and therapeutic applications.

Similarly, CAR-T cells, engineered to target specific antigens expressed on tumour cells, can be identified and characterised based on the expression of CAR molecules and activation markers. Common markers used for CAR-T cell analysis includeCD3, CD4, CD8, and CAR-specific markers such as CD19 or CD20 (9). By assessing the expression levels of these markers, researchers can evaluate CAR-T cell functionality, persistence, and anti-tumour activity, guiding the development and optimisation of CAR-T cell therapies.

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Applications of
Cellular Phenotyping

The applications of cellular phenotyping extend across various fields, including immunology, oncology, stem cell research, and infectious diseases. In immunology, cellular phenotyping is used to study immune cell subsets, activation states, and functional responses to pathogens or therapeutic interventions (10). In oncology, it aids in the identification of tumour-infiltrating immune cells, assessment of immune evasion mechanisms, and monitoring of immune checkpoint blockade therapies.

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References

[1] BD FACSuiteTM Application | BD Biosciences [Internet]. www.bdbiosciences.com. [cited 2024 May 14]. Available from: https://www.bdbiosciences.com/en-gb/products/software/instrument-software/bd-facsuite-application#Overview
[2] Maecker HT, McCoy JP, Nussenblatt R. Standardizing immunophenotyping for the Human Immunology Project. Nature Reviews Immunology [Internet]. 2012 Feb 17;12(3):191–200.  
[3] Perfetto, S. P., Ambrozak, D., Nguyen, R., Chattopadhyay, P., & Roederer, M. (2011). Quality assurance for polychromatic flow cytometry using a suite of calibration beads. Nature Protocols, 6(12), 1-17.
[4].Omana-Zapata I, Mutschmann C, Schmitz JL, Gibson S, Judge K, Monika Aruda Indig, et al. Accurate and reproducible enumeration of T-, B-, and NK lymphocytes using the BD FACSLyric 10-color system: A multisite clinical evaluation. 2019 Jan 28;14(1):e0211207–7.
[5] Mousset CM, Hobo W, Woestenenk R, Preijers F, Dolstra H, van der Waart AB. Comprehensive Phenotyping of T Cells Using Flow Cytometry. Cytometry Part A. 2019 Feb 4;
[6] Velounias RL, Tull TJ. Human B-cell subset identification and changes in inflammatory diseases. Clinical and experimental immunology. 2022 Dec 1;210(3):201–16.  
[7] Del Zotto G, Antonini F, Pesce S, Moretta F, Moretta L, Marcenaro E. Comprehensive Phenotyping of Human PB NK Cells by Flow Cytometry. Cytometry Part A. 2020 Mar 21.  
[8] Banik A, Prabhakar S, Kalra J, Anand A. An enriched population of CD45, CD34 and CD117 stem cells in human umbilical cord blood for potential therapeutic regenerative strategies. Current Neurovascular Research. 2014;11(4):312–20.  
[9]Tantalo DG, Oliver AJ, Scheidt B von, Harrison AJ, Mueller SN, Kershaw MH, et al. Understanding T cell phenotype for the design of effective chimeric antigen receptor T cell therapies. Journal for ImmunoTherapy of Cancer. 2021 May ;9(5):e002555.
[10].Mishra HK. Clinical Applications of Flow Cytometry in Cancer Immunotherapies: From Diagnosis to Treatments. Methods in Molecular Biology. 2022 Dec 14;93–112.