Olink Proteomics Sample Operation Specification

In the discovery of disease biomarkers, analysis of immune responses, and mechanism research, the quantitative precision and throughput of Olink proteomics have continuously improved. Although mass spectrometry has advantages in depth, it faces certain challenges in clinical translation and application scenarios with large sample sizes or rare samples. In this context, Olink® Target and Explore platformsstand out, thanks to theirextremely high sensitivity, low sample requirement, and good batch consistency, becoming one of the mainstream technologies in proteomics research in recent years. However, the high performance of Olink technology is based onstandardized sample handling. Any pre-treatment errors may cause quantitative bias or affect data interpretation. Therefore, mastering and executing strict sample handling protocols is a key step to ensure project success.

 

I. Introduction to Olink Proteomics Technology

Olink employs Proximity Extension Assay (PEA) technology, which combines antibody specificity with nucleic acid amplification to achieve high sensitivity and high specificity in protein quantification. Its features include:

1. Extremely low sample volume requirement (~1μL)

2、No need for sample denaturation or enrichment

3. High throughput (capable of detecting 92~3000 proteins simultaneously)

4. NPX (Normalized Protein eXpression) data output, comparable across batches

 

Due to its reliance on antibody affinity and amplification efficiency, Olink proteomics is extremely sensitive to sample conditions (such as hemolysis, degradation, type of anticoagulant), making detailed operational specifications particularly important.

 

II. Olink Proteomics Sample Types and Collection Requirements

Supported sample types:

1. Plasma: Preferred, recommended to be collected using EDTA anticoagulant

2. Serum: Also suitable for most panels, but expression of certain proteins may have deviations

3. Cell culture supernatant: Usable but requires removal of particles and filtration

4. Tissue lysate, cerebrospinal fluid, urine: Supported by some panels, confirmation required in advance

 

Recommended collection protocol:

  

Project	Requirements explanation
Type of collection tube	K2EDTA purple tube (preferred), avoid heparin (interference with amplification)
Operation time	Complete centrifugation within 30 minutes after blood collection to avoid interference from cell lysis
Centrifugation conditions	2000xg, 10 minutes, 4°C
Plasma aliquoting	≥30μL per tube, avoid multiple freeze-thaw cycles
Storage temperature	Long-term storage at -80°C, storage at <-20°C is not recommended for more than 1 week

  

Biotech company Biotech ParkIt is recommended to use low protein adsorption tubes to avoid sample adhesion loss.

 

III. Olink Proteomics Sample Quality Control and Troubleshooting Suggestions

To ensure data quality, the following issues need to be focused on during the early stages of Olink sample preparation:

1. Avoid hemolysis

Free hemoglobin in hemolyzed samples can interfere with optical readings and nucleic acid amplification, it is recommended to:

• Carefully inspect the sample color (pink to red is abnormal)

• Set a Hemolysis Score threshold, and exclude if necessary

 

2. Protein degradation control

Protease activity or repeated freeze-thaw cycles can lead to loss of target proteins, it is recommended to:

• Add protease inhibitor cocktail (for tissue or cell lysates)

• Strictly control the number of freeze-thaw cycles,in principle ≤1 time

 

3. Cross-contamination and inter-batch variation

• All sample operations should be carried out in a clean environment, using filter tips

• Sample information management should be systematic, it is recommended to useunique identification + cryogenic barcode

 

Given the precious nature of samples and the high testing costs on the Olink platform,it is recommended to first conduct a Pilot Study (e.g., 10-20 cases)to perform preliminary evaluations on panel selection, sample stability, and intergroup differences. Biotree Biotech can assist with end-to-end support from experimental design to bioinformatics analysis. Olink proteomics technology offers a revolutionary solution for high-throughput, low-sample-volume protein quantification research. However, its high dependence on sample condition means that preprocessing must be impeccable. By executing scientific and standardized sample handling protocols, researchers can maximize the accuracy of the data and the power of biological interpretation.Biotree Biotechpossesses extensive project experience and a rigorous sample quality control system. Whether you are in the early exploration phase of a project or in the stage of clinical cohort research, we can providecomprehensive solutions for sample management, platform testing, and data interpretation support.。

 

Biotree Biotech - A high-quality service provider for bioproduct characterization and multi-omics mass spectrometry detection.

 

Related services:

Olink Proteomics