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Olink Precision Proteomics Analysis Service
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Olink Precision Proteomics Analysis Service

Product Description
Case Analysis
Result display
Sample delivery
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Product Description

With the accomplishment of human whole genome sequencing, medical research has moved from genomics to the post-genomic era. The research value of proteins as functional executors of life activities is gaining more and more attention. Protein biomarkers can serve as important features in pathophysiology, bridge the gap between genome and phenotype, and have a profound impact on improving future healthcare, especially in precision medicine. However, the progress has been stunted by the lack of technologies that can provide high specificity, sensitivity, and throughput. The emergence of Olink technology, however, can break through these obstacles and accelerate the development of proteomics in the field of precision medicine.

Technology principle

Olink protein detection platform is based on the neighbor extension technology Proximity Extension Assay (PEA) for protein detection. For each protein to be detected, a pair of antibodies is designed at two adjacent antigenic epitopes, which are coupled to a specific DNA single strand with paired bases at the end of the single strand. When the pair of antibodies binds to the target protein, the two adjacent DNA single strands are hybridized and complemented and extended in an enzymatic reaction, which then forms a double-stranded template. The detection of the double-stranded template using microfluidic qPCR or NGS enables the quantitative analysis of the protein. This technique cleverly converts protein quantification to DNA quantification, solving the problem that proteins cannot be amplified, thus enabling the detection of trace amounts of proteins.

Technical advantages

1. High specificity: High specificity by using two antibodies to recognize one protein, labeling and full validation by Barcode.

2. Wide dynamic range: The entire panel spans 10 log values, allowing simultaneous consideration of proteins of different abundance.

3. High sensitivity: The detection sensitivity is as low as fg/mL level, which can detect thousands of disease-related low abundance proteins (especially low abundance biomarkers in plasma and serum samples) at the histological level.

4. High detection throughput: 96/384/3072panel, each panel can detect 88 samples at a time. By combining qPCR and NGS high throughput detection platform, it can produce millions of protein data per week.

5. Micro sample: 48-384 protein markers can be detected simultaneously in 1μL plasma, and more than 3000 protein markers can be detected in 8μL plasma.

6. High reproducibility: strict quality control, 99.8% of proteins without cross-reactivity, mature detection platform, high data quality, good reproducibility, suitable for large data analysis requirements, and the reproducibility requirements of clinical translational applications.

Application areas

Biomarker research: disease risk assessment, disease typing, disease prediction, disease diagnosis and prognosis.

Drug discovery: drug target discovery, clinical treatment effect evaluation, basic research.

Case Studies

Case 1: Next generation plasma proteome profiling for monitoring health and disease with Olink technology

Article Title: Next generation plasma proteome profiling to monitor health and disease

Publication date: May 2021

Journal: Nature Communications (IF=14.9, Q1)

Sample type: plasma

Technology used: Olink proteomics technology (Olink Explore 1536 / Olink Target 96)

Article Overview

In this paper, a NGS-based Olink Explore 1536-plex assay was introduced and compared with a qPCR-based Olink Target 96 assay to confirm the consistency of the assay results. The stability of protein differences was also confirmed by correlation analysis of plasma protein profiles with the whole genome. The plasma protein profiles of healthy and type 2 diabetes (T2D) cohorts were analyzed to identify relevant protein markers and confirmed to correlate with clinical assays for identifying T2D high-risk groups and stratifying metformin treatment response in T2D patients to provide dosing guidance.

 

 

 

Case 2: Single-cell transcriptomics combined with interstitial fluid proteomics can clarify cell type-specific immune regulation in atopic dermatitis

Artical Title: Single-cell transcriptomics combined with interstitial fluid proteomics defines cell type-specific immune regulation in atopic dermatitis

Publication date: April 2020

Journal: The Journal of allergy and clinical immunology (IF=10.79, Q1)

Sample type: cutaneous blister

Technology used: Olink proteomics technologies (inflammation, immune response, neurology panels), single-cell transcriptome

Article Overview.

Atopic dermatitis (AD) is the most common chronic inflammatory skin disease, affecting up to 20% of children and 7%-10% of adults, with the degree of impact varying by population. Both defective skin barrier and abnormal immune activation are specific features of the disease, but their interactions remain unclear. Inflammation centered on TH2 is one of the key known causative factors, as demonstrated by the clinical efficacy of dupilumab. Dupilumab is a recently approved IL-4 receptor a blocker for the treatment of moderate to severe AD. However, only 30% of AD patients treated with dupilumab had a 90% reduction in eczema area and severity index. The average disease improvement rate for broadly acting immunosuppressive agents (e.g., cyclosporine platform) is only about 70%, suggesting that pathways outside the known immune axis must be relevant in causing the disease. Importantly, animal models only reflect a few aspects of AD, and therefore, further knowledge and understanding of AD through studies on human tissues is essential.

The authors characterized AD at both transcriptomic and proteomic levels. A characteristic upregulation of cytokines typical of AD such as IL13 and IL22 was found in TH2 and TH22 cells, respectively, compared to healthy controls. However, these mediators were also found in proliferative T cells and natural killer T cells, which also express the antimicrobial cytokine IL26. Overall, it was not T cells, but myeloid cells that were most strongly enriched in AD.

The authors found the most significantly upregulated proteins in AD blister fluid proteomic analysis included dendritic cells (CLEC7A, double-regulated proteins, EREG) and macrophage products (CCL13). These data suggest that the use of aspiration blistering techniques offers several advantages over conventional biopsies, including better transcriptome resolution of skin cells. This technique, combined with proteomic information from interstitial fluid, was able to reveal novel inflammatory factors that shape the cellular and proteomic microenvironment of AD.

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The upper figure shows the distribution of NPX values for all samples in the selected panel as a box plot, where blue represents samples with sample QC pass.

IQR质控图

The upper figure shows samples with no significant outlier distribution in the presentation of the upper dot plot, no abnormalities in the degree of data dispersion within any sample were found.

 

The upper panel presents protein expression in all samples, where each column represents a sample and each row represents a species protein, and the gradient color shows the relative expression of proteins in the samples.

 

The upper panel presents the correlation between all proteins, with the horizontal and vertical axes representing each protein, the names are shown at the bottom and right, and the gradient color shows the pearson correlation coefficient values between samples.

Notices of samples delivery

Olink Sample Handling Guidelines

Serum collection

1. collect whole blood into a serum collection tube.

2. Allow the blood to clot at room temperature, this usually takes 15-30 minutes. In samples left for more than 60 minutes, it is possible that clotted cells may begin to lyse.

3. centrifugation at 1000-2000g for 10 minutes in a refrigerated centrifuge (2-8°C) to remove the blood clot.

4. immediate transfer of serum to clean tubes, dispensing 50-200 μL per tube.

5. Store the dispensed sample at -80°C.

Plasma collection.

1. Collect whole blood into anticoagulant-treated collection tubes (EDTA, sodium citrate or heparin);

2. centrifuge at 1000-2000g for 10 minutes in a refrigerated centrifuge (2-8°C) to remove cells from the plasma.

3. immediately transfer the plasma into clean tubes, dispensing 50-200 μL per tube.

4. Store at -80°C.

Collection tube type and size.

Any collection tube used for the collection of serum or plasma is acceptable.

Please keep all samples collected in the same way and in the same collection tube (same brand, same size, etc.).

Transport materials.

Samples should be sent in temperature-resistant, non-protein-bound plastic, such as a 96-well PCR plate, preferably one with a full skirt edge, and a sealing film that is resistant to temperatures as low as -80°C and of high quality.

Sample volume.

Volume of each sample ≥ 40 μL. recommended volumes: 96-well plates, 50 μL/sample; Eppendorf tubes, 100-500 μL/sample.

Sample quantity.

For analyses performed by Olink, only one tube of plasma or serum per subject/patient is enough.

Sample labeling.

Clearly mark the sample plate or tube with a temperature-resistant label or marker, e.g., a simple alphanumeric code for later identification ("A, B, C", "1, 2, 3" or "A1, A2, A3").

Transport temperature.

Samples should normally be shipped on dry ice (they should still be frozen on arrival). Please contact your local courier company for detailed information and local regulations regarding risk assessment and the use of dry ice for human sample transport.

Note: For handling of other types of samples, such as tissue and cell lysates, fine needle biopsies, exosomes, microdialysis fluids, bronchoalveolar lavage, cell culture media, dried blood spots, synovial fluid, cerebrospinal fluid, plaque extracts, urine, saliva, etc., please contact company salesperson to obtain.

Sample Requirements.

Sample Type     

Storage Media

Sample size requirements 

Transport method

Serum      Freezing tube

100~500μL/sample

Dry ice transport

Serum 96-well plate

50μL/sample

Dry ice transport

Plasma Freezing tube

100~500μL/sample

Dry ice transport

Plasma

96-well plate     

50μL/sample

Dry ice transport

 

Lead time

The standard process from sample receiving to data delivery has a lead time of approximately 2 months (40 business days). Customized analytical requirements need additional assessment of service period.

Note: Serum/plasma samples should be processed within 1 hour of collection in accordance with the Olink Sample Processing Guidelines. Processing other types of samples are customized service, please contact sales staff.

Technical support services:

1. Completing the analysis according to the contract.

2. Submitting documents for the project closure report with a description of the project results.

3. Providing documents of the project closure report, off-line raw data and related bioinformatics analysis results generated during the analysis process.

4. Providing free consulting services within 6 months of data delivery.

5. The final data can be retained for another 3 months after delivery. A fee assessment is required to extend the data retention period.

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