Olink Proteomics: Blood/Serum Sample Handling Protocol
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Carefully inspecting the color of the sample (pink to red indicates abnormal hemolysis).
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Setting a hemolysis score threshold and discarding samples as necessary.
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Add a protease inhibitor cocktail (for tissue or cell lysates)
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Limit freeze-thaw cycles to a maximum of one cycle
In the fields of disease biomarker discovery, immune response analysis, and mechanistic research, the quantitative accuracy and throughput requirements of Olink proteomics have been steadily increasing. While mass spectrometry offers significant advantages in terms of depth of analysis, it faces challenges in clinical translation, particularly when dealing with large sample volumes or rare sample types. In this context, the Olink® Target and Explore platforms have emerged as leading technologies in recent proteomics studies, thanks to their exceptional sensitivity, minimal sample requirements, and excellent batch-to-batch consistency. However, the high performance of Olink technology depends on standardized sample handling protocols. Any errors during sample preprocessing may lead to quantitative discrepancies or undermine data interpretation. Therefore, adhering to stringent sample handling protocols is essential to ensure the success of the project.
Introduction to Olink Proteomics Technology
Olink employs the Proximity Extension Assay (PEA) technology, which combines antibody specificity with nucleic acid amplification to achieve highly sensitive and specific protein quantification. Key features include:
1. Extremely low sample volume requirements (~1 μL)
2. No need for sample denaturation or enrichment
3. High throughput (enabling simultaneous detection of 92 to 3000 proteins)
4. NPX (Normalized Protein eXpression) data output, ensuring comparability across batches
Due to its reliance on antibody affinity and amplification efficiency, Olink proteomics is highly sensitive to sample conditions such as hemolysis, degradation, and anticoagulant type. Therefore, establishing detailed operational protocols is crucial to ensure reliable results.
Olink Proteomics Sample Types and Collection Requirements
1. Supported Sample Types
(1) Plasma: Preferred. EDTA anticoagulant is recommended for collection.
(2) Serum: Usable for most panels, though some proteins may show biased expression.
(3) Cell Culture Supernatant: Usable, but particles must be removed through filtration.
(4) Tissue Lysates, Cerebrospinal Fluid, Urine: Supported by certain panels, but prior confirmation is required.
2. Recommended Collection Protocol
MtoZ Biolabs recommends using low-protein-binding tubes to prevent sample adhesion loss.
Olink Proteomics Sample Quality Control and Troubleshooting Guidelines
To ensure high-quality data, the following issues should be addressed in the preprocessing of Olink samples:
1. Avoid Hemolysis
Hemolyzed samples contain free hemoglobin, which can interfere with optical readings and nucleic acid amplification. We recommend:
2. Control Protein Degradation
Protease activity or repeated freeze-thaw cycles can lead to the loss of target proteins. It is advised to:
3. Prevent Cross-Contamination and Batch Variability
All sample processing should be carried out in a clean environment using filter tips. Sample management should be systematized, and it is recommended to use unique identifiers and storage barcodes.
Considering the high value of Olink samples and the cost of detection, we recommend conducting a Pilot Study (e.g., 10-20 samples) to evaluate panel selection, sample stability, and inter-group variability. MtoZ Biolabs provides one-stop support, from experimental design to bioinformatics analysis. Olink proteomics offers a revolutionary solution for high-throughput, low-sample-volume protein quantification. However, its reliance on sample integrity means that preprocessing must be performed with precision. By following scientifically rigorous sample handling protocols, researchers can maximize the accuracy and biological relevance of their data. MtoZ Biolabs has extensive project experience and a robust sample quality control system. Whether you are in the exploratory phase or conducting clinical cohort studies, we can provide comprehensive support for sample management, platform testing, and data interpretation.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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