T and B lymphocytes are principal components of the host immune system. Based on outstanding expertise and rich experience, Creative Biolabs has developed a panel of assays to assess and screen the immune effects of cancer epitopes determined by the biological activity of T and B cells. Our featured assay services are listed as follows.

Fig.1 The roles of adaptive T cells and B cells in anti-tumor immunity.¹

✔ T cell-based Biological Assays

Helper T cells play a core role in adaptive immune immunity, as they modulate multiple immune cell responses to pathogens. They are required in B cell activation to produce antibodies, and induced T cells secrete perforin and granzyme to kill tumor cells. Creative Biolabs offers the following T cell-based biological assays for investigating the cellular mechanism of action.

• Activation

Flow cytometry is used to detect T cell activation by activation markers of CD69 and CD25, in combination with T cell-specific markers, such as CD3, CD4, and CD8.

• Proliferation

Creative Biolabs provides the following assays for the evaluation of T cell division after antigen stimulation:

✔ CFSE
✔ ³H-thymidine
✔ BrdU-based Methods

• Cytotoxicity

The assessment of the target cell death induced by antigen-specific T cells is a direct way to identify the functional cancer peptides. The specific killing of target cells can be measured by the standard assays, such as:

✔ Radioactivity Chromium (⁵¹Cr) Assay
✔ Non-radioactivity LDH Release Assay

• Degranulation

Degranulation is quantitatively measured by detecting the expression of lysosome-associated membrane protein 1, LAMP-1/ CD107a.

• T Cell Help

T cells are activated when they recognize the antigen peptides presented by MHC molecular on B cells and help activation of cognate B cells. Creative Biolabs offers services monitoring the T cell-mediated B cell activation by co-culturing B cells with T cells that are activated by specific antigens and quantitatively measuring the B cells' responses, including:

✔ Antibody Production
✔ Cytokines Secretion in the Culture Supernatant

✔ B cell-based Biological Assays

Investigation of pathogen-induced B cell responses and the subsequent production of antibodies from activated B cells is a valuable method for screening antigenic peptides of cancer. Creative Biolabs offers comprehensive assays based on the assessment of B-cell activity and function pathways to profile antigen-specific b cell responses in whole blood and PBMCs.

• Activation

B cell activation marker CD69 combined with B cell-specific markers, such as CD19, CD20, CD22, and CD27, can be used to detect activated B cells by flow cytometry.

• Proliferation

By labeling the cells with fluorescent proliferation dye and measuring changes in the fluorescent intensity by flow cytometry.

• Antibody Production

ELISA is applied to detect the presence of antibodies and their concentration in serum, plasma, or culture supernatant.

• Neutralization

The neutralization assay is performed in two forms:

✔ Detecting the influence of antibodies on antigen-expressing cells' viability, proliferation, and infectivity.
✔ Measuring the binding affinity of the antibody to the ligand.

• Cytotoxicity

Antibody-dependent cellular cytotoxicity (ADCC) assay and complement-dependent cytotoxicity assay are both available. NK cells or other engineered cells stably expressing FcγRIIIa and luciferase are used as effector cells. Multiple in vitro and in vivo analysis methods are offered, such as target cell lysis, effector cell activation, and real-time monitoring of effector cells and target cells.

• Hemagglutination Inhibition

The hemagglutination assay and the hemagglutination inhibition assay are constructed to measure the antibody's specific binding to the HA molecule of the virus.

Creative Biolabs offers a full range of cancer epitope discovery assays to in-depth validate your potential candidates at an early stage. Our experienced scientists will participate in the whole process of your program to accelerate your project development. Please contact us by email for detailed information.

Reference

1. Briquez, P. S.; et al. Engineering Targeting Materials for Therapeutic Cancer Vaccines. Frontiers in bioengineering and biotechnology. 2020, 8, 19.

For Research Use Only | Not For Clinical Use