Technology

Proximity Extension Assay (PEA) technology

The latest breakthrough in biomarker discovery is the Proximity Extension Assay (PEA) technology developed by Olink Proteomics (Uppsala, Sweden). Proximity Extension Assay (PEA) is a 96-plex immunoassay for high throughput detection of protein biomarkers in body fluids. For each biomarker, a matched pair of antibodies linked to unique oligonucleotides binds to the respective protein target. When placed in close proximity, the stretch of nucleotides hybridizes and the annealing product is then amplified by PCR and detected in multiplexed fashion in a high throughput fluidic chip system. Specificity and reproducibility of the assays are rigorously quality controlled (Assarsson et al., PLoSOne 2014) (see Fig.1).

Fig. 1 Olink Proximity Extension Assay (PEA)
Fig. 1 Olink Proximity Extension Assay (PEA) workflow. Bottom right side is the plate layout: 88 samples (each spiked with four internal controls: 2x Incubation, Extension and Detection), 2x External sample control (plasma pool used to calculate Intra- and Inter-CVs), 3x Negative control (used to calculate the limit of detection) and 3x Interplate control (IPC) (used for normalization).

 

HMGU hosts the first Core Facility certified for Olink technology in Germany. Using Proximity Extension Assay (Olink Proteomics), we provide accurate quantification of up to 1164 human clinically relevant protein biomarkers in plasma and other biological samples using only 1µL sample per panel. Each panel of 92 biomarkers is focused on a specific area of disease or key biology process, for a total of 15 different panels: six disease area-focus panels, e.g. cardiovascular disease (CVD II and CVD III), oncology (ONCOLOGY II and ONCOLOGY III), NEUROLOGY, INFLAMMATION, IMMUNO-ONCOLOGY and seven additional panels focused on important biological processes such as CARDIOMETABOLIC, METABOLISM, CELL REGULATION, DEVELOPMENT, IMMUNE-RESPONSE, NEURO-EXPLORATORY and ORGAN DAMAGE. Additionally, MOUSE EXPLORATORY is designed for exploratory mouse studies and offers 92 assays for murine proteins which are also detectable in human serum and plasma.

Fig.2 Dynamic range of selected Olink panels.
Fig.2 Dynamic range of selected Olink panels.

 

Olink panels span a broad dynamic range of protein concentrations and they are designed to cover expected ranges in normal and pathological samples. This extremely sensitive antibody-based technology allows for accurate quantification of human plasma proteins present in concentrations below picogram per milliliter, such as interleukines and hormones (see Fig.2). As only very little volume of body fluid is necessary, this methodology can be applied for various epidemiologic or clinical research cohorts.

Literature: Assarsson E, Lundberg M, Holmquist G, Björkesten J, Thorsen SB, Ekman D, Eriksson A, Rennel Dickens E, Ohlsson S, Edfeldt G, Andersson AC, Lindstedt P, Stenvang J, Gullberg M, Fredriksson S. Homogenous 96-plex PEA immunoassay exhibiting high sensitivity, specificity, and excellent scalability. PLoS One. 2014 Apr 22;9(4):e95192.

 

ANALYSIS SERVICE WORKFLOW:
The Core Facility Proteomics accepts samples from HMGU and from external users (upon collaboration agreement between your institution and HMGU).  We offer you support in any step of the analysis service, from study design, sample submission to data analysis. Please see below the detailed workflow to run your samples on our Olink platform:

 

Please download the Sample Submission Sheet and the Project Proposal here. Carefully read the section “Sample preparation and shipment terms” in the Project Proposal!