Our kits are instrumental in quantifying the cell-mediated immune responses induced by new vaccine candidates, providing critical data for evaluating their protective efficacy against infectious diseases like influenza, HIV, malaria, or emerging viral threats. This includes assessing antigen-specific T cell activation and cytokine production.
As a beacon of innovation in immunology, Creative Biolabs provides bespoke enzyme-linked immunosorbent spot (ELISpot) based kit development services, and we provide a wide range of ELISA kits for the specific detection of disease-related proteins. We leverage our unparalleled expertise to help clients unlock precise, single-cell insights into immune responses, driving forward critical advancements.
How Does It Work?
The ELISpot assay, a highly sensitive modification of the conventional sandwich ELISA, offers a unique window into the functional activity of individual immune cells. Initially developed for detecting antigen-specific antibody-secreting B cells, its application has broadened significantly. The core principle involves coating a microplate with a specific capture antibody. When immune cells, such as T cells or B cells, are added and stimulated, any secreted analytes (e.g., cytokines, chemokines, or antibodies) are immediately captured by the immobilized antibody. Subsequent detection with a biotinylated secondary antibody, followed by an enzyme-conjugated streptavidin and a chromogenic substrate, generates visible spots. Each spot precisely corresponds to a single activated cell secreting the target molecule, providing a direct, quantitative, and qualitative measure of cellular immune responses. This visualization capability makes ELISpot exceptionally valuable for accurately detecting rare antigen-specific T or B cells within heterogeneous cell populations like PBMCs.
Fig.1 Illustration of the ELISpot assay.Distributed under CC BY-SA 3.0, from Wiki, without modification.
Advantages of Our ELISpot Platform
Creative Biolabs is dedicated to developing highly sensitive, flexible, quantitative, and user-friendly ELISpot based kits. This advanced method offers several significant advantages for detailed immune monitoring:
Single-Cell Resolution: ELISpot permits direct visualization of the secretory product(s) from individual activated or responding cells, providing unprecedented detail that bulk assays cannot match.
Qualitative Information: Beyond mere presence, the assay yields specific information regarding the type of cytokine or other immune molecule secreted, defining the immune response profile.
Quantitative Frequency Analysis: It accurately quantifies the frequency of responding cells within a test population, allowing for precise determination of antigen-specific cellular frequencies.
Exquisite Sensitivity: ELISpot's remarkable sensitivity enables the detection and frequency analysis of rare antigen-specific cells, a feat that was often impossible to perform with earlier immunological techniques, thereby opening new avenues for research and diagnosis.
Our ELISpot Based Kit Development Services
At Creative Biolabs, our ELISpot based kit development service is a comprehensive, client-centric offering designed to meet your precise research and diagnostic needs. We specialize in providing fully customized ELISpot solutions, moving far beyond generic off-the-shelf options. Our service spans the entire development lifecycle, from initial consultation and strategic assay design to meticulous antibody development, rigorous assay optimization, and extensive validation. We leverage our deep immunological expertise to select the most appropriate capture and detection antibody pairs, optimize cell stimulation conditions, and fine-tune every parameter to achieve unparalleled sensitivity and specificity for your target analytes. Whether you require a single-analyte kit or a multiplexing solution (e.g., FluoroSpot), we ensure your custom kit delivers robust, reproducible, and reliable results, perfectly integrated into your experimental workflow.
Applications
Vaccine Efficacy Assessment
Immunotherapy Monitoring in Oncology
In the rapidly evolving field of cancer immunotherapy, ELISpot kits enable precise monitoring of anti-tumor immune responses. This includes tracking the activity of immune cells (e.g., cytotoxic T lymphocytes) in response to checkpoint inhibitors, CAR-T cell therapies, or oncolytic viruses, offering insights into treatment efficacy and patient response.
Autoimmune Disease Research
For conditions such as rheumatoid arthritis, multiple sclerosis, or lupus, our custom ELISpot kits facilitate the identification and characterization of autoreactive T cells or B cells. This helps researchers understand disease pathogenesis and evaluate the impact of immunomodulatory treatments on specific cellular responses.
Transplantation Immunology & Rejection Monitoring
Post-transplant, the ELISpot assays are crucial for detecting alloreactive T cells that mediate graft rejection or contribute to graft-versus-host disease (GVHD). This allows for timely intervention and personalized immunosuppressive therapy adjustments, significantly improving patient outcomes.
Infectious Disease Diagnostics & Surveillance
Beyond tuberculosis, our custom kits can be developed for the specific detection of cell-mediated immunity to a wide array of pathogens, assisting in the diagnosis of latent infections, monitoring disease progression, or assessing population-level immune status following outbreaks.
Published Data
1. Enzyme-Linked Immunosorbent Spot Assay for the Detection of Specific T Cells
Fig.2 ELISpot assay of two representative cases of CD8+ T cells isolated from PBMCs.1,3
In this work, researchers evaluated Wilms' tumor 1 antigen-specific cytotoxic T lymphocytes (WT1-CTLs) using the ELISpot assay and compared it with MHC tetramer analysis for the first time as a validation method in DC-based cancer immunotherapy. The ELISpot assay, conducted on peripheral blood mononuclear cells, demonstrated consistent reproducibility, daily reliability, and a linear response. WT1-specific immune responses were observed in 73.9% of patients who received DC vaccinations. The correlation analysis showed ELISpot assay had a very strong correlation (Spearman`s ρ, p =0.82) with WT1 tetramer analysis. These results confirm that the ELISpot assay is a reliable and validated tool for monitoring immune responses in patients undergoing DC vaccination targeting WT1.
2. Enzyme-Linked Immunosorbent Spot Assay-Based Evaluation of Cytokines
Fig.3 Representative image of ELISpot plates.2,3
The goal of this research was to use the ELISpot assay to measure PBMCs secreting IL-10, TNF-α, IL-1β, and IL-17A cytokines in early and established Rheumatoid arthritis (RA) patients and healthy controls. The ELISpot assessed cytokine expression in freshly isolated PBMCs from whole blood to show that IL-17A-secreting PBMCs were markedly increased in early RA patients. The study demonstrates that the ELISpot assay is a reliable tool for evaluating cytokine secretion in immune responses, as it can measure in-vitro cytokine secretion by PBMCs, quantifying the number of cytokine-producing cells and detect even a single cell that secretes the cytokine of interest among millions. This method provides valuable insights into cytokine profiles and their association with RA disease activity.
Service Highlights
Tailored Solutions
Fully customized kit components and protocols are provided, optimized precisely for your unique antigens, cell types, and target analytes, ensuring maximum relevance and performance.
Rigorous Quality Control
Every kit undergoes comprehensive validation for sensitivity, specificity, reproducibility, and stability. This guarantees consistent and reliable results batch after batch, with extensive optimization of all components and protocols for robust performance.
Scalability & Manufacturing
Flexible manufacturing capabilities support projects from small-scale pilot batches to large-volume production, ensuring a seamless transition for your research or clinical needs.
Dedicated Scientific Support
Our commitment extends beyond kit delivery; ongoing expert scientific and technical support is available, ensuring you maximize the utility and success of your custom ELISpot assay.
Q&A
-
Q: Why should I choose a custom ELISpot kit over a commercially available, off-the-shelf option?
A: While commercial kits offer convenience, a custom ELISpot kit developed by Creative Biolabs is specifically optimized for your unique research needs, including novel antigens, specific cell populations, or particular target analytes. This tailored approach ensures unparalleled sensitivity, specificity, and performance that generic kits, designed for broad applications, simply cannot match. It guarantees that the assay precisely reflects your experimental parameters, leading to more accurate and relevant data.
-
Q: What types of analytes can your custom ELISpot kits detect?
A: Our development capabilities are extensive. We can create custom ELISpot kits to detect a wide range of secreted analytes, primarily cytokines (e.g., IFN-γ, IL-2, IL-4, IL-10, TNF-α), chemokines, granzymes, perforin, and even antigen-specific antibodies secreted by B cells. If you have a specific secreted protein in mind, our scientific team can assess its feasibility for ELISpot detection and optimize the assay accordingly.
-
Q: What is the typical sample type required for your ELISpot assays?
A: ELISpot assays primarily utilize live immune cells that are capable of secreting the target analyte upon stimulation. The most common sample type is peripheral blood mononuclear cells (PBMCs), isolated from whole blood. However, depending on your research needs, we can also optimize kits for use with dissociated tissue samples, lymph node cells, splenocytes, or other isolated immune cell populations.
-
Q: Can your ELISpot kits be developed for multiplexed detection?
A: Absolutely. We have extensive experience in developing multiplexed ELISpot kits that allow for the simultaneous detection of two or more analytes from the same single cell. This capability provides richer, more nuanced data on cellular immune responses. Our team will work with you to design and optimize the fluorescent detection systems and imaging protocols necessary for successful multiplexing.
-
Q: Can you help with the selection of appropriate antigens or stimuli for my ELISpot assay?
A: Yes, antigen and stimulus selection are critical steps for a successful ELISpot assay. Our scientific team, with years of experience, can provide expert consultation on selecting or designing the most appropriate antigens, peptides, or mitogens to stimulate your target immune cells effectively and elicit the specific responses you aim to measure.
Cytokine Bead Arrays (CBA) and conventional ELISA assays can provide extremely useful information in certain contexts but lack the sensitivity and accuracy of ELISPOT for the detection and enumeration of rare antigen-specific cells. Creative Biolabs is a world leader in the development of ELISA based kits and now offers ELISpot based kits for specific detection of disease-related proteins. If you are interested in our ELISA kits development, please feel free to contact us for more details.
References
- Higuchi, Yumiko, et al. "Enzyme-linked immunosorbent spot assay for the detection of Wilms' tumor 1-specific T cells induced by dendritic cell vaccination." Biomedicines 3.4 (2015): 304-315.
- Dissanayake, Keerthie, et al. "Potential applicability of cytokines as biomarkers of disease activity in rheumatoid arthritis: Enzyme-linked immunosorbent spot assay-based evaluation of TNF-α, IL-1β, IL-10 and IL-17A." PloS one 16.1 (2021): e0246111.
- Distributed under Open Access license CC BY 4.0, without modification.
For Research Use Only.
