ELISA Assay Automation Important Considerations
We recently finished our Ask the Expert discussion on “How to get the most out of automating your ELISA-like assays, dos and don’ts.” During this Ask the Expert session, we covered topics related to ELISA assay automation and ELISA-like assay automation, including assay transfer, optimization, validation, challenges, benefits and specific questions regarding data generation.
This Ask the Expert session was hosted by Phillip Gorman. Philip is the senior researcher for the High Throughput Antibody Discovery team at Boehringer Ingelheim Pharmaceuticals Inc. He obtained his BS in Physiology and Neurobiology from the University of Connecticut and an MS degree in Biology from Southern Connecticut State University. He has held many roles within the Pharmaceutical and Biotechnology industries with focus on application development and support for research in genetics, immunotoxicology and immunology. Within the last 17 years, Philip has gained knowledge and experience in assay development, automation and automation integration. Since 2011, he has been expanding the use of automation for antibody discovery within BI, focusing on ELISA, AlphaLISA and MSD assay formats.
Below is a sneak peek of the discussion, for a full transcript, please see – Ask the Expert – How to get the most out of automating your ELISA-like assays, dos and don’ts.
How long does it take to get an automated system up and running? Months, weeks?
Well, the answer to that question would depend greatly on the size and complexity of the automated system. If you are referring to something simple like a plate stacker attached to a plate reader, it would say hours. As you get into larger and more complex systems then the time needed to get them running and validated would increase. There are several considerations to be mindful of. For example, if you are integrating a piece of equipment that has never been integrated with the controller software that you are using, you might discover some problems that can extend the installation and validation timelines. It has been my experience with the small-to-medium sized, fully automated platforms that I have built that it can easily take a couple weeks to get everything running smoothly. Larger, more complex systems can take a few months to set-up and validate.
What did you find was the biggest benefit in automating your assay process? What was the biggest challenge?
In my opinion, there are 2 big benefits, consistency in your assay process and time management of the scientists. Running assays on a properly designed and validated system provides a level of consistency that is very difficult to duplicate with manual pipetting. In an automated environment you know that every plate was handled exactly the same way as every other plate so the data, even over long periods of time, are directly comparable. Automating assay work also allows scientists to focus their attention on non-routine experiments or tasks while an assay is running in the background, effectively acting as a ‘force multiplier’.
The biggest challenge that I have experienced, especially when working with novice users, is trying to change the way some scientists think about assay development and validation in the context of automated systems. If you work with automation regularly, you have to be able to think about resource allocation within an automated system as it pertains to your assay workflow. One small change in process in an automated workflow can significantly affect the time it takes to complete. Changing reagent addition steps, or which component will do which steps requires a type of fluid thinking that can be difficult for some scientists to grasp.
I am wondering how the assay transfer works. How difficult is it to transfer and how do you validate after automation?
The term “assay transfer”, in this context, simply means moving an assay from the bench (manually completed) to being run on some form of automation. This assumes that the assay has already been properly developed and validated, if appropriate. Ideally, when performing an assay transfer to an automated platform you would compare the values of your control samples, be it EC/IC50, Signal-to-Noise ratio or some other quantitative measure. They are run in a manual assay and in an automated assay. If everything has been done correctly those values should be equivalent, within a reasonable %CV.