In Vitro CAR-T Cell Functional Analysis with Peptide-DC Vaccination

All products and services are For Research Use Only and CANNOT be used in the treatment or diagnosis of disease.

pMHC-directed CAR-T cells, combined with a peptide-DC vaccine, represent a promising approach towards enhancing anti-tumor immune responses. At Creative Biolabs, we are committed to providing high-quality services and support to researchers developing novel immunotherapy approaches. Our team of experts is dedicated to providing in vitro functional analysis services to help researchers evaluate the efficacy and potency of peptide-DC vaccine in boosting pMHC-directed CAR-T cells. With our cutting-edge technologies and expertise, we offer a comprehensive range of assays including cytotoxicity assays, proliferation assays, cytokine production assays, and activation marker analysis, among others.

Fig.1 Schematic of functional parameters on activated CAR T cells.¹

In Vitro CAR T Immunotherapy Assays

Our in vitro CAR-T cell functional analysis services provide a comprehensive characterization of CAR-T cells co-cultured with peptide-loaded cell lines. Our experienced team will perform a series of assays to evaluate the key functional properties of CAR-T cells, including cytotoxicity, cytokine secretion, proliferation, and persistence. We will provide detailed experimental protocols, perform the experiments with precision and efficiency, and deliver clear and reliable data analysis. Some common methods used for the functional analysis of CAR-T cells include:

Proliferation Assay

The proliferation ability of CAR-T cells is measured by various methods after the CAR-T cells are cultured with peptide-loaded cell lines. We utilize techniques such as flow cytometry, BrdU incorporation assays, cell counting, and real-time imaging to accurately assess CAR-T cell proliferation. CAR-T cells are labeled with a proliferation dye (such as CFSE or CellTrace Violet) and co-cultured with target cells. The dilution of the dye in daughter cells over time is used as a measure of cell proliferation.

Phenotype Assay

Our phenotype assay service offers an investigation into the influence of peptide-DCs on CAR-T cell phenotype. This service enables a comprehensive analysis of the cellular characteristics and markers expressed by CAR-T cells when exposed to peptide-DCs. Through this assay, we aim to provide valuable insights into the impact of peptide-DC interactions on the phenotype of CAR-T cells.

Cytokine Assay

CAR-T cytokine assay is important for evaluating the potency and functionality of CAR-T cells, as well as for monitoring immune responses and potential adverse effects. Our service can be customized to detect specific cytokines of interest, such as IL-2, TNF-α, and IFN-γ, and can be performed using different techniques, such as ELISA, or flow cytometry. The results of the assay can provide valuable information for optimizing CAR-T cell therapy and improving patient outcomes.

Cytotoxicity Assay

Our team of scientists and technicians will perform the cytotoxicity assay using various methods, including chromium release assay, flow cytometry-based assay, and LDH release assay.

Our experienced team of scientists is able to customize our in vitro and in vivo functional assays to meet the specific needs of your research or drug development project. Please contact us to learn more about our in vitro assay service and how it can advance your scientific goals.

Reference

Si, Xiaohui, et al. "Preclinical evaluation of CAR T cell function: in vitro and in vivo models." International Journal of Molecular Sciences 23.6 (2022): 3154.