Transparent validation for data you can trust (Olink Explore)
Quality, rigor and transparency are very important values for Olink Proteomics. Consequently, all of our assays are rigorously quality controlled and our validation data is made freely available. Below you can see the details of exactly how we validate each individual assay, and the product as a whole.
The variables described below are presented for each assay on the individual biomarker pages for Olink® Explore 1536. Please note that the NPX values presented in the validation data show NPX above background. Background is defined as the median of negative control measurements and used to define the expected background level (zero).
Validation data overview
You can download an Excel file containing all available validation data for the assays in Olink Explore 1536:
DOWNLOAD VALIDATION DATA
Limit of Detection (LOD)
In vitro sensitivity (calibrator) curves using recombinant antigen are determined in multiplex format, for each of the 384-plex panels within Olink Explore. Note that in some cases, no suitable antigen is available and no calibrator data is presented. Limit of detection (LOD) is defined as 3 standard deviations above background and reported in pg/mL. The y-axis shows NPX above background, which is defined as the median of negative control measurements and used to define the expected background level (zero).
Example calibrator curve from assay validation
High dose hook effect
The high dose hook effect is seen when there is an antigen excess relative to the reagent antibodies, resulting in falsely low results. In such cases, a significantly lower value may lead to erroneous interpretation of results.
Therefore, the hook threshold is determined for each analyte and reported in pg/mL and this value is presented on each individual biomarker page.
The analytical measuring range is defined by the lower limit of quantification (LLOQ) and upper limit of quantification (ULOQ) and reported in pg/mL. Quantification limits of LLOQ and ULOQ are calculated using relative error <30% and CV <30%.
Example analytical measuring range data from assay validation
The levels of protein measured in a number of commercial plasma samples are presented in sample distribution plots. Healthy subjects (n=24) and samples obtained from patients with a range of diseases (n=48). These include inflammatory, cardiovascular (n=12) , autoimmune (n=12) & neurological diseases (n=12), as well as cancer (n=12). These data provide a general idea of the NPX range to expect but cannot cover all potential levels in clinical samples. The y-axis shows NPX above background, which is defined as the median of negative control measurements and used to define the expected background level (zero).
Example sample distribution plot from assay validation
Intra-assay variation (within-run) is calculated as the mean CV for 6 individual samples, within each of 7 separate runs during the validation studies. Inter-assay variation (between-runs) is calculated as the mean CV, for the same 6 individual samples, among 7 separate runs during the validation studies.
Across all 1472 assays, the mean intra-assay and inter-assay variations observed were 8% and 11%, respectively.
Inter-site variation (between-site) was also investigated during the validation of Olink Explore at three sites (two service laboratory sites and Olink R&D), to estimate the expected variations in values between different laboratories running the same samples, with different operators using different equipment. Each site was trained and instructed to perform the analysis according to the same routine. Each site performed 6-7 independent runs.
The analysis of results from these multi-site studies are currently being finalized and will be updated here as soon as they are available.