ELISA Assay Development
Typically at the beginning of an ELISA project, our scientists will discuss with clients for assay formats and reagent availability. Clients can provide antibodies or antigens, or we identify and obtain potential antibodies and antigens specific for the target. A typical assay development process includes the following steps:
- Characterize the specificity of the antibodies and antigens using Western blot.
- Determine optimal dilutions for all antibodies and antigens
- Determine the throughput: Assays can be developed in 96w or 384w format
- Optimize incubation times, temperatures and buffers
- Optimize the assay for maximum sensitivity and precision
- Determine lower and upper limits of detection
- Optimize standards and standard curves for quantitation
We have a suite of automation equipment allowing us to run ELISA assays in full screening mode in either 96w or 384w formats. We also possess a team of scientists who have extensive experience in development and implementation of drug screening assays.
Once the assay is established, a typical workflow for screening includes the following:
- Coating plates, using Multidrop or Combi
- Prepare dilutions of test articles, such as antibodies and lysates, using Agilent Bravo liquid handling station
- Transfer test articles to assay plates using Agilent Bravo
- Incubate and wash using BioTek plate washer
- Add secondary antibody, then detection buffer
- Read on EnVision or SpectraMax
- Data analysis with scatter plots, hit rates, Z’ factors, etc.