Ask a Quanterix Scientist: The Inside Scoop on Simoa® Evaluation Kits
By Danielle Fontaine, PhD
Quanterix Field Applications Scientist
Simoa® Advantage kits including NfL, pTau-181 and Neuro 4-Plex E (Aß40, Aß42, GFAP, NfL) kits come with everything you need to run an assay. However, if you are new to working with an assay and need to perform validation experiments before using these kits, you likely need additional materials to complete these validation studies. This is where Evaluation Kits come in – these kits are available for select Advantage kits and contain enough calibrator diluent, sample diluent, and calibrator concentrate any time you need additional materials. Evaluation kits are lot-matched to the assay kits to ensure the most reliable results and should be ordered at the same time as the assay kits. Read on as we explore frequently asked questions relating to the use of evaluation kits.
What types of experiments require additional material?
When performing initial validation experiments, extra material may be necessary. Evaluation kits provide reagents for experiments such as spike/recovery or dilution/recovery assays. The calibrator concentrate can also be used to generate custom controls for use across your experiments, or if you have panels to test and you need spiked samples. One other example is for investigators who are validating the lower limit of quantification (LLOQ) for a new assay and need to generate their own serial dilutions of the calibrator.
What is included in each Evaluation Kit?
Each Evaluation kit contains the reagents listed below, though the volume of each reagent will vary depending on the specific assay. Assay Kit instructions that contain details on what volumes are provided will be included with every kit.
- Calibrator concentrate: Each Evaluation kit will include a Calibrator that is more concentrated than the highest calibrator concentration provided in the assay kits. The Calibrator concentrate can be diluted as appropriate. If you have ordered a multiplex assay and its corresponding Evaluation kit, separate Calibrator Concentrates will be provided for each analyte. The Certificate of Analysis (which can be searched for here) provides information on the calibrator concentrations within a lot, expected control values, and provides the lot numbers for each reagent within a kit.
- Calibrator diluent: This reagent will help you create your own serial dilutions of the provided calibrator concentrate.
- Sample diluent: This dilution reagent can be used to optimize sample dilutions and create panels or dilution linearity studies.
Why is the diluent different between calibrators and samples?
This is a great question! The behavior of antigens in a laboratory made buffer may be different than in its natural matrix, like plasma or serum. To take this into consideration, we have developed separate diluents for calibrators and samples so we can better align the behavior of the antigen to the two sample matrices.
Why do the Evaluation kits need to be lot-matched to the assay kits?
There are biological components that are used to create each set of calibrators that may have some variability between lots. By ordering Evaluation kits that are lot-matched with the assay kits you plan to run your sample on, the calibrator and sample diluents are better to your eventual experimental results.
Do all assays have evaluation kits?
No, not all kits have lot-matched evaluation kits available. However, we may have separate (but lot-matched!) calibrator concentrate that may be able to help run validation studies. Please contact Quanterix to discuss available options.
Have a question about evaluation kits that was not addressed here? You can find more information on our Evaluation kits within the Assay Technical Notes section of the Customer Portal. For assistance with experimental design for assay performance evaluation or for any other questions regarding Simoa® assays, please contact TechSupport@Quanterix.com
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