T Cell-based High Throughput Multiplexed Assay

Creative Biolabs is expertise in the mechanism of T cell antigens recognition and is experienced in applying immunological technologies for antigen discovery. Now, we provide high-throughput multiplexed assays to identify T-cell epitopes for basic and translational research, such as drugs and vaccine design for infectious diseases.

Current High-throughput Multiplexed Approaches

Multiple approaches are utilized to develop a high throughput multiplexed method to explore a large scale of potential epitopes in the antigen protein. Most of the approaches display the peptide-MHC complex on cells, such as tumor cells, yeast, or baculovirus, and co-culture with cognate T cells or use soluble TCRs to assess T cell binding and activation. Recently, a team delivered the genome-wide antigen to cells and processed it by endogenous MHC-I molecules to imitate the natural antigen-presented pathway and use the granzyme B activity as the effector molecular to monitor T-cell responses.

Fig.1 Discovery of T cell antigens by high-throughput screening of synthetic minigene libraries.¹

High Throughput Multiplexed Assay Services at Creative Biolabs Services

At Creative Biolabs, we provide a systematic high-throughput antigen genome screening assay to identify cancer epitopes associated with T cell immunity, providing potential targets for diagnostics, antibody-mediated therapy, and vaccine development.

Workflow

• Cancer Antigen Proteome Library Construction

A method that comprehensively profiles cancer antigens is crucial for identifying T cell epitopes that are physiologically meaningful in vivo. We have developed a high-throughput technology to construct an ORF library covering the entire coding domain of cancer antigens. In detail, each OFR is cloned into our vector and expressed using in vitro transcription/translation kit.

• Polyclonal T-cell Stimulation

PBMCs are stimulated with full-length antigens to produce enough antigen-specific T cells. The specific T cells are cultured to more than 10 billion, and CD4+, CD8+ T cells are sorted for the following screening.

• Epitope Screening

Expanded CD4+ T cells are co-cultured with the synthesized antigen protein library and cognate antigen-presenting cells (APC).

Expanded CD8+ T cells are mixed with antigen proteins presented by artificial APCs that are generated from HLA cDNA and peptides co-transfected COS-7 cells.

• Monitor T Cell Immune Response

A read-out assay, such as a ³H-thymidine incorporation assay or an ELISpot assay for cytokine secretion, is adopted to screen the antigen-specific responses.

Fig.2 Screening results of IFNγ-release ELISpot.¹

Analytes Available

We provide the following popular cytokine and chemokine release measurement services to monitor T cell responses.

Related High-throughput Techniques

• T and B Cell-based ELISA Assay
• T Cell-based Cytometric Bead Array Assay

Creative Biolabs provides high-quality services with enhanced productivity, freeing up your time for other core work at the time. Our experts are 24h online for consultants. Please contact us to discuss your demands.

Reference

1. Hondowicz, B. D.; et al. Discovery of T cell antigens by high-throughput screening of synthetic minigene libraries. PloS one. 2012, 7(1): e29949.

For Research Use Only | Not For Clinical Use