Hi everyone,
I am doing ELISA for plasma and serum samples as a part of my Master's project. I used to use ELISA kits that involve high and low quality control samples that helped me to ensure that I am doing the ELISA properly and get valid results. I am planning to do ELISA from other companies, too. However, they do not involve any quality control samples. Is it necessary to use quality control samples for ELISA? How can I ensure that if I did the ELISA I am going to get valid results in this case? How do I prepare quality control samples?
Hi Manar, many companies produce commercial ELISA kits for the same goal. presence of quality control samples supplied by kits standardized your results based on their manufacturing process so accurate results get occurred. using other quality control samples not supplied by kit will give you a new standard for result calculation that deviate from kit manufacture but in this case all readings under the same error factors. so you can not produce good ELISA without standardize all ELISA components
regards
1. You can purchase control samples from other company.
2. You can do yourself control samples and evaluate it.
3. You can purchase and use standarts from national systems of standarts likes NIBSC.
It's dependent from your analit - its cost, its stability etc.
Hi Manar,
Like others said it's depending on what you are looking for in your ELISA. Is it from tissue sample or plasma/serum sample.
Just as an example of what you can do. Recently we have analysed a lot of insulin on elisa plate. It was plasma sample from coming from different center. We have choose to use lyphocheck from bio-rad as a control. The purpose of the standard was just to ensure that nothing wrong had happens for each elisa. We haven't use it as an interplate calibrator.
Hi Manar,
Quality Controls are very important in validation of your results. You can use known positive samples as a postive control and known negative samples as negative control.Before that you should set up standard OD for each QCs.
Thank you for all your replies. I am doing standards and samples in duplicates to ensure the precision of my pipetting. I want to control for accuracy; i.e to which level I am closer to the true value of the analyte in the sample. The analytes I am testing are proteins in serum or plasma such as resistin and adiponectin. I believe that even if I prepared my own controls, I am not going to validate my ELISA, since the manufacture standards where the ELISA kits are tested are different from what I am going to standardize in my lab.
Dear Helena,
what is recombinant protein? I mean what is it made of? How is it going to be useful for ELISA validation? much appreciated for further explanation.
The way to validate an ELISA (or any analytical assay) is described best at the ICH Q2_R1:
The way to validate an ELISA is used CLSI EP-12 guide and quality planning.
Greetings
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