Sandwich ELISA

The Sandwich Enzyme-Linked ImmunoSorbent Assay (ELISA) is a sensitive and robust method which measures the antigen concentration in an unknown sample. The antigen of interest is quantified between two layers of antibodies: the capture and the detection antibody. These antibodies must bind to non-overlapping epitopes on the antigen. Either monoclonal or affinity-purified polyclonal antibodies can be used as capture and detection antibodies, depending on cost, the dynamic range and the sensitivity of the final assay.

 Please CLICK HERE and describe your ELISA assay development needs so that we can provide a quote or set up a call.

The general protocol is as follows:

1. Apply a sample of known antigen to a surface, often in the well of a microtiter plate. The antigen is fixed to the surface to render it immobile.
2. The plate wells or other surface are then coated with blocking buffer.
3. Detecting antibody, usually diluted in blocking buffer, is applied to the plate for binding to the antigen coated on the plate.
4. The plate is washed, so that unbound antibody is removed. After this wash, only the antibody-antigen complexes remain attached to the well.
5. The second antibodies, which will bind to any antigen-antibody complexes, are added to the wells. These second antibodies are coupled to the substrate-modifying enzyme.
6. Wash the plate, so that excess unbound antibodies are removed.
7. Apply a substrate which is converted by the enzyme to elicit a chromogenic or fluorescent signal.
8. View/quantify the result using a spectrophotometer or other optical device.


GenWay offers custom Sandwich ELISA development. Our experienced R&D team can work with our clients for determining factors such as cost, sensitivity, specificity and accuracy of each assay.  Please CLICK HERE and describe your ELISA assay development needs so that we can provide a quote or set up a call.

The Sandwich ELISA can be mainly applied for the following experimentation or research:

  • Fast and accurate detection of antigen concentration in an unknown sample
  • High detection sensitivity and reproducibility are required

Sandwich ELISA development typically takes 2 months to complete, depending on the availability of the antigen and the antibodies, which can be provided by the customers or developed by GenWay. For each project, in addition to the assay,

GenWay delivers the protocol and all the necessary reagents. Post-delivery customer support is available.
Service prices, detailed specifications, and further information are provided upon request by contacting