Development of ELISpot
With the wide application of enzyme-linked immunoassay technology in the medical and biological fields, based on the basic principle of ELISA technology, enzyme-linked immune absorbent spot (ELISpot) was established. As a biotechnology researcher representing the international level, Creative Diagnostics can provide professional ELISpot development services according to your research needs.
Why Choose Development of ELISpot?
ELISpot is an extension and new development of quantitative ELISA technology. When you cannot find a suitable ELISpot kit on the market to detect your target, when you need to develop a new quantitative analysis method for a certain antibody or cytokine, you can choose our ELISpot development services.
Questions to Consider Before Choosing This Service
- Do you need to develop cell-based immunoassays?
- Do you need an immunoassay method that can accurately reflect the level of antibodies and cytokine?
- Is the cytokine or antibody you need to detect readily metabolized in body fluids or bound to target organs?
If your answer is yes, then this development service can basically meet your ELISA needs.
Advantages and Disadvantages of ELISpot
In ELISpot, coated primary antibodies are used to capture cytokines secreted by cells in culture. Then, biotin-labeled secondary antibody/fluorescein-labeled secondary antibody, enzyme-labeled avidin, and BCIP/NBT substrate were added in sequence. The detection results were displayed in the form of enzyme-linked spot coloration.
The Main Advantages of ELISpot
- Functional assays of living cells based on the single-cell level.
- Compared with the traditional ELISA method, the sensitivity is greatly improved.
- ELISpot has no complicated in vitro cell expansion process, is simple and economical, and can be used for high-throughput screening.
The Main Disadvantages of ELISpot
- Incomplete removal of cells disrupts spot formation.
- The preparation of the ELISpot plate and the culture of cells need to be carried out under sterile conditions, and there are restrictions on the site.
Fig. 1 The Flowchart of ELISpot
Development of ELISpot
ELISpot measures secretion from individual cells, not the average level of a population of cells. In the development of ELISpot, the complication lies in the sterile culture of live cells and the stimulation of antigens. We will strictly follow industry standards and relevant laws to implement the workflow of ELISpot development services.
After our ELISpot development team believes that this development service can meet your testing needs, we will evaluate your testing needs and design a solution based on them.
Cell culture is a critical step in ELISpot. Based on rich experience and superb technology, we can cultivate cells that are in good condition, have high vitality, and function well.
In ELISpot, antigens are used to stimulate cultured cells to produce corresponding cytokines. Our high-end technology platform supports us to prepare enough high-purity antigens that meet your experimental needs.
We need to develop monoclonal primary and secondary antibodies for ELISpot. Due to the process involved in cell culture, the requirements for monoclonal antibodies are much higher than for capturing antibodies in conventional ELISA. We will prepare endotoxin-free, high-affinity antibodies for you.
To ensure that the antibody binds specifically to the antigen after the antibody is synthesized, we need to analyze the level of biomolecular interaction of the antibody.
In the development of ELISpot, we usually label the secondary antibody with biotin, so that the biotin can be combined with the enzyme-labeled avidin for chemical enzyme-linked color development. We can also use fluorescein to label the secondary antibody, eliminating the subsequent chemical development step, and can detect multiple cytokines simultaneously.
The kit components used in the development of ELISpot must be sterile, and for the high price of plates, we usually choose PVDF plates with better adsorption for pre-coating of primary antibodies. After the development materials are selected, the optimized ELISpot program development is carried out.
In this development step, we will repeatedly optimize the antibody dilution, background noise, sensitivity, etc. to provide you with a high-quality ELISpot development solution that meets your needs in all aspects.
In this step, we will use your designated biological samples to verify the developed ELISpot to ensure the correctness of the development strategy.
After the ELISpot development protocol has been validated, we will communicate with you to produce the specified ELISpot kit containing all reagents and components according to your requirements.
Result/Material Delivery
Development Protocol
All Experi-mental Data
Ready-to-use ELISA Kits
Validated Assay Results
Creative Diagnostics' development of ELISpot, tailored to your needs, provides the most powerful tools for your immunology research. At the same time, we also provide corresponding ELISA testing services and also provide comprehensive solutions for your ELISA kit customization needs. If you are interested in our ELISpot development services, please feel free to contact us.
