T Cell-based ³H-thymidine Assay

Screening cancer epitopes, particularly neoepitope antigens, has become increasingly important in drug discovery and immunotherapy in recent years. Creative Biolabs provides various T and B cell-based epitope analysis assays, including T cell-based 3H-thymidine assay, to analyze cancer pathogenic epitopes.

Cancer Epitope and T Cell-based 3H-thymidine Assay

Cancer antigen epitopes are specific molecules that target T and B cell receptors and induce complex immune responses. Cancer epitopes are typically derived from somatic mutations that cause tumorigenesis and may progress to various diseases and cancer. T cell proliferation analysis is a reliable and reproducible method to determine the presence and frequency of epitope-specific T cells. The use of a ³H-thymidine incorporation assay to investigate T cell proliferative response is a validated tool to screen and test cancer pathogenic epitopes.

Fig.1 Schematic representation of ³H-thymidine assay.¹

³H-thymidine Assay-Based Epitope Analysis Services at Creative Biolabs

Creative Biolabs has developed a robust T cell assay platform to provide reliable ³H-thymidine assay to our clients worldwide. Based on your objectives, our experts will design scientific experiments and conduct high-quality studies to support your research.

Workflow of Mouse Model-based Cancer Epitope Discovery Using ³H-thymidine Assay

Fig.2 The workflow of T cell-based ³H-thymidine assay. (Creative Biolabs)

Induce Epitope-specific T Cell Response

T cell proliferation detects antigen-responsive T cells generated from in vitro and in vivo models.

T Cell Proliferation Analysis

• T cells cultures are incubated with ³H-thymidine for several hours to days.
• As a result, dividing cells incorporated the ³H-labeled thymidine into the replicated DNA strand during the co-culturing period.
• Afterward, the radioactive is measured using a scintillation counter.

Data Interpretation

The data interpretation is a necessary aspect of the assay. We have established standardized protocols to identify a cut-off value to distinguish positive results from negative ones.

Fig.3 Cell proliferation to human thyroglobulin peptides.²

Highlights of Our T Cell-based ³H-thymidine Assay

• Simple and easy to perform
• Relatively high throughput
• Test large numbers of antigens/peptides

Creative Biolabs is a professional provider of bioassays. Our years of program development services provided us with a wealth of experience, allowing us to collaborate closely with our clients to provide T cell assays, including design and operation. Our assay provides clients with consistent and reproducible results. Please feel free to contact us us to get more information.

References

1. Di Blasi, D.; et al. Guidelines for low-frequency antigen-specific T cell results analysis: Dye-based proliferation assay vs 3H-thymidine incorporation. Journal of immunological methods. 2020, 487: 112907.
2. Flynn, J. C.; et al. Pathogenic human thyroglobulin peptides in HLA-DR3 transgenic mouse model of autoimmune thyroiditis. Cellular immunology. 2004, 229(2): 79–85.

For Research Use Only | Not For Clinical Use