How to Make a Top-Quality Immunoassay

Have you ever wondered how to develop a top-quality immunoassay for a fraction of the cost of what you can buy from an assay company? If you have the time, you can develop similar assays for just the cost of the materials. What assay companies provide, and what you’re paying for, is the one-time development cost, material cost, and the quality control of each batch.

What about all those specifications that are already configured with a commercial assay; ones such as lower limit of quantitation, accuracy, precision, dilutional linearity and others? There are great resources available to help you make an assay that has the same high quality as you expect from a commercial supplier, explain all those terms, and tell you how to develop a robust assay.

Here are some helpful resources for immunoassay development:

  • NIH Chemical Genomics CenterImmunoassay Methods. This website provides general guidelines for development, optimization and validation of immunoassays. This is where we suggest you start. Following the guidelines on this site will likely increase the chances that you will be able to develop a reproducible immunoassay that can be used in your research. This is your cookbook style tool that will take you step-by-step through the process of developing an immunoassay.
  • FDA Guidance for IndustryBioanalytical Method Validation. The FDA released this document in 2001 and it remains the general guidance for bioanalytical assays. This is guidance for industry involved in developing drugs and other regulated products, and the concepts apply to all laboratories not just pharmaceutical development labs.
  • European Medicines AgencyGuideline on bioanalytical method validation. This document is a fantastic resource for assay parameter definitions. Its intent is to provide recommendations for bioanalytical methods for measuring biologic drug concentrations in clinical samples. The assay requirements set forth may be more stringent than that required by a research laboratory.
  • In order to develop an immunoassay using Luminex® xMAP® Technology, we have a resource page with protocols. The only difference between developing on Luminex technology is the substrate to which you attach a capture antibody or other capture molecule.

We’ve provided five resources for developing your own immunoassays and there are definitely more available. These should cover the vast majority of the knowledge needed to develop a high-quality immunoassay.

What is your experience developing your own immunoassays? What are other good resources?