T and B Cell-based ELISPOT Assay

Antigen-specific humoral and cellular immunity can be identified by T and B cell activation responses. Creative Biolabs provides the advanced Enzyme-Linked Immunospot (ELISpot) assay to have an insight into antigenic epitopes by measuring cytokines secretion activity.

Enzyme-linked Immune Absorbent Spot (ELISpot) Assay for Cancer Epitope Analysis

Investigation of antigenic peptide-specific humoral and cellular immunity can be monitored by detection of cell activation response, such as cell expansion, cell-specific killing, and cytokine and other effector molecules production. By measuring cytokine concentration changes in PBMC, we can directly validate T cell stimulation by specific antigen peptides. Due to the accurate, sensitive, reproducible results of ELISpot, the assay became the gold standard for evaluating antigen-specific immune response since it was developed and is widely used for various studies, including CTL monitoring, viral antigen-related cell response diagnosis in clinical experiments, and vaccine candidates.

Fig.1 The schematic of ELISpot assay.¹

ELISPOT Assay Services at Creative Biolabs

ELISpot is typically used to evaluate antigen-specific immune responses of T and B cells. This assay is also reliable in studying cancer epitopes that can elicit host immune responses. With the mature and formal platform developed by our in-house talented experts and technicians, Creative Biolabs offers highly sensitive T and B cell-based ELISpot assay for cancer epitope analysis for our clients.

Workflow

The ELISpot assay is ideal for measuring both immunoglobulins secreted epitope-specific B cells and the cytokines release of specific epitope-stimulated T cells. The procedure contains the following main steps:

Fig.2 The experiment procedure of ELISpot assay. (Creative Biolabs)

ELISpot Assay Validation

During assay establishment, the ELISpot assay is optimized and validated to assess the fitness of the trial and ensure the optimum performance for your PBMCs or spleen cell samples.

• Start Cell Numbers
• Specific Antigen Concentration
• Antibody Concentrations
• Image System

Available ELISpot Assays

Compared to other methods measuring individual cytokine-secreting cells, the ELISpot assay always outranks others due to its unique sensitivity to detect low-frequency antigen-specific lymphocytes. ELISpot can also be used to assess multiple cytokines produced by T-helper (Th) cells, Th2 cells, Th17 cells, and B cells.

The popular ELISpot assay includes but is not limited to below.

Data Analysis

• Automatic Report. Color spots are imaged and analyzed using an automated system to interpret the results to quantitate immune response.
• Optimized two-color ELISpot. Two capture antibodies conjugated with different color reactions are used to discriminate the cell populations producing different cytokines. And the total number of spots and subpopulation statistics for multiple cytokines are automatically reported.

Key Features and Advantages

• High Sensitivity

Allows the detection of rare antigen-specific lymphocytes and figures out one individual cytokine production cell out of 105 cells in culture.

• High-throughput

Many samples in a trial can be simultaneously analyzed for immune kinetics and ease of operation.

• Quantitative Measurement

This technique has a wide quantitative range in measuring cytokines production.

Creative Biolabs provides reliable ELISpot-generated data that is core in the specific immune responses in cancer epitope identification studies. Our experts are available to discuss your research and help you design the most effective application of the assay. Please contact us and tell us your needs.

Reference

1. Navarrete, MA. ELISpot and DC-ELISpot Assay to Measure Frequency of Antigen-Specific IFNγ-Secreting Cells. Methods Mol Biol. 2015, 1318: 79-86.

For Research Use Only | Not For Clinical Use