Development of Sandwich ELISA

Development of Sandwich ELISA

As a leading ELISA manufacturer, Creative Diagnostics provides customers with tens of thousands of sandwich ELISA kits for a variety of targets. On this basis, we use our many years of ELISA research experience to provide customers with sandwich ELISA development services. We can analyze, design, develop, and optimize sandwich ELISA according to customers' needs, which can significantly shorten customers' experimental time.

Why Choose Development of Sandwich ELISA?

When you are studying an antigen/antibody and need to quantitatively detect its content in the sample, but there is no ready-made ELISA kit on the market that meets the requirements, or you are developing higher sensitivity and wider analytical range when looking for a wide range of new sandwich ELISA assays, you can choose our custom sandwich ELISA development.

Questions to Consider Before Choosing This Service

  1. Is your test substance a macromolecular antigen or antibody?
  2. Does the target you need to detect have two or more binding sites?

If your answer is yes, then basically you can choose the custom sandwich ELISA development. If you still have doubts, we will provide you with detailed consulting services, and discuss together to choose the ELISA development service that best meets your needs.

Advantages and Disadvantages of Sandwich ELISA

Sandwich ELISA is divided into double-antibody sandwich ELISA and double-antigen sandwich ELISA.

In the double-antibody sandwich ELISA, the antigen is bound to different sites by two antibodies, the capture antibody, and the detection antibody. The capture antibody is immobilized on an ELISA microplate, and the detection antibody undergoes a color reaction by binding to the antigen. This method is suitable for the detection of macromolecular antigens with multiple sites.

In the double-antigen sandwich ELISA, the antigen is immobilized on the ELISA microplate. After the antibody in the sample to be tested is combined with the antigen, the enzyme-labeled antigen is added and combined with the antibody to carry out a color reaction. This method is suitable for detecting antibodies.

The Main Advantages of Sandwich ELISA

  • Two antibodies/antigens are used for higher sensitivity.
  • Sandwich ELISA can be detected by direct or indirect methods, with high flexibility.
  • Two antibodies/antigens are used and bind to different sites of the target, making them highly specific.

The Main Disadvantages of Sandwich ELISA

  • Double antibody sandwich ELISAs are highly demanding on antibodies and may require custom antibody development.
  • Double-antigen sandwich ELISA requires the preparation of enzyme-labeled antigens according to the antigen structure, and the operation is complicated.

The Flowchart of Sandwich ELISA - Creative DiagnosticsFig. 1 The Flowchart of Sandwich ELISA

Development of Sandwich ELISA

In order to provide customers with the best quality custom sandwich development service, our entire research and development process will be strictly implemented in accordance with the following contents.

After evaluating your sandwich ELISA development needs and determining that this service can meet your research needs, our expert team will provide you with a detailed and highly feasible sandwich ELISA development plan according to your requirements.

High-purity antigens are the cornerstone of successful ELISA development. The double-antibody sandwich method only involves the preparation of one antigen, but the double-antigen sandwich method involves the synthesis of two antigens. We will prepare the antigens strictly according to the structure of the antibody to be tested.

Both the double-antibody sandwich method and the double-antigen sandwich method involve the preparation of primary antibodies. We have extensive experience in antibody preparation and can certainly provide you with a tailored primary antibody.

After antibody synthesis and antibody conjugation, we will use antibody affinity assay technology to analyze the level of biomolecular interactions to ensure successful protocol development.

The double-antibody sandwich involves antibody pairing, and this step is the key to the development of sandwich ELISA. We perform at least 90-pair matching tests to find the best matching antibody pair. It should be noted that the binding site of the detection antibody to the target protein needs to be different from the site to which the capture antibody binds. Commonly used antibody pairing methods are as follows:

  • The capture antibody is a monoclonal antibody, and the detection antibody is a polyclonal antibody.
  • The capture antibody is a monoclonal antibody, and the detection antibody is a monoclonal antibody, and the two types of monoclonal antibodies target different epitopes.
  • The capture antibody is a polyclonal antibody, and the detection antibody is a polyclonal antibody, and the two polyclonal antibodies are generally derived from different hosts.

In the double-antibody sandwich, the secondary antibody needs to be labeled. We generally introduce the biotin-avidin system and select the corresponding activated biotin and reaction conditions according to the types of labelable groups (amino groups, aldehyde groups, sulfhydryl groups, etc.) and molecular acidity that the antibody carries. After biotin labeling, it does not affect the biological activity of the labeled substance.

The key to the double-antigen sandwich is the preparation of the enzyme-labeled antigen, and we will select the most suitable label for it according to the structure of the antigen.

Next, we select all the reagents (including assay diluents, conjugate stabilizers, substrates, stop solution, etc.) and materials involved in sandwich ELISA to develop sandwich ELISA.

In this step, we will optimize the sandwich ELISA for issues such as non-specific binding of antibodies, high substrate concentration, poor reproducibility, etc.

After optimizing the sandwich ELISA development protocol, we will use the samples you specify for assay validation to ensure that the protocol fully meets your requirements.

After verifying that the sandwich ELISA, we will produce the sandwich ELISA kit according to your requirements, and provide you with a ready-to-use sandwich ELISA kit including all reagents and components.

Result/Material Delivery

Development Protocol

Development Protocol

All Experi-mental Data

All Experi-mental Data

Ready-to-use ELISA Kits

Ready-to-use ELISA Kits

Validated Assay Results

Validated Assay Results

Creative Diagnostics looks forward to providing you with new or optimized sandwich ELISA development services. For more details on the development of sandwich ELISA, please contact us. It has to be said that we also provide reliable ELISA testing services and provide comprehensive solutions for your ELISA kit customization needs.

The service is for research only, not for clinical use.
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