ELISA Kit Development

Enzyme-linked immunosorbent assay (ELISA) is a method of adsorbing a known antigen or antibody on a solid-phase carrier, adsorbing the antibody or antigen in the sample to be tested by specific binding of the antigen-antibody, and adding enzyme-labeled antibody and substrate. Under the action of the corresponding enzyme substrate, a colored substance is generated, and the color depth is proportional to the content of the corresponding antibody or antigen in the marker, so that the content of the antigen or antibody can be determined. ELISA has a simple operation process and can quickly analyze a large number of parallel samples, which is of great significance in the diagnosis of pathogenic infections.

There are four common ELISA types: direct ELISA, indirect ELISA, sandwich ELISA and competitive ELISA.

Direct ELISA	Indirect ELISA
The antigen is immobilized on the ELISA plate, followed by direct detection of the antigen with an enzyme-labeled antibody.	The antigen is bound to the ELISA plate and the detection antibody is added to bind specifically to the antigen, followed by the addition of the enzyme-labeled secondary antibody for detection and color development using the substrate.
Advantages Short operation procedure, few experimental steps No need to use secondary antibody to avoid cross-reactivity Fewer experimental steps, less prone to errors	Advantages Enzyme secondary antibodies can enhance the signal and improve sensitivity Greater flexibility, same secondary antibody can be applied to many different primary antibodies No direct labeling of primary antibodies to retain the most immunoreactivity
Disadvantages Direct enzyme labeling of primary antibodies is required, which is relatively costly Poor flexibility, requires specific primary antibodies for each target protein The detection sensitivity is not high enough: no secondary antibody is used, and the signal is not amplified	Disadvantages Cross-reactivity may exist Longer experimental cycle than direct ELISA
Sandwich ELISA	Competitive ELISA
The capture antibody is bound to the ELISA plate, the antigen is immobilized by the capture antibody, and then detected by direct ELISA or indirect ELISA.	Antigens or antibodies in the sample compete to bind specific amounts of enzyme-labeled antibodies or antigens. The higher the concentration of antigen in the sample, the weaker the output signal, and the signal output is inversely proportional to the amount of antigen in the sample.
Advantages High specificity, using capture and detection antibodies that are both specific to the test substance Suitable for complex samples, antigens can be used for detection without purification Greater flexibility and sensitivity, either by direct or indirect methods	Advantages Good reproducibility: less variation between replicate samples and analyses High flexibility: based on direct, indirect or sandwich ELISA More robust: less sensitive to sample dilution and sample matrix effects than sandwich ELISA
Disadvantages Not suitable for detection of small molecules High requirement for paired antibodies	Disadvantages Cumbersome operation

ELISA Kit Development

With the spread of various infectious animal diseases all over the world, it has brought a huge impact on the social economy. In order to control the epidemic of infectious animal diseases, it is necessary to develop a rapid and high-throughput screening method for animal diseases. ELISA is a powerful assay that is widely used for large-scale screening of animal pathogens. Currently, the demand for inexpensive and reliable custom ELISA kits is growing rapidly. At BioVenic, we can customize a range of ELISA kits for animal pathogen detection based on your needs. If you have any needs, please feel free to contact us, we can provide you with high-quality products and services.

The Workflow of ELISA Kit Development

There are many things to consider when developing an ELISA kit. We can design different ELISA kits for you to capture and detect the target antigen depending on your detection target. The following is the workflow of ELISA kit development.

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