Olink Proteomics: Blood/Serum Sample Handling Protocol

1. Sample Types

Olink panels have undergone rigorous validation with plasma and serum samples. Other extensive sample types, such as cerebrospinal fluid, tissue lysates, and cell supernatants, have also been successfully analyzed using Olink.

2. Sample Collection and Pre-Analytical Handling

Pre-analytical Sample Processing

Standardization of the sample preparation procedure is crucial. Throughout the process, from study design and sample collection to data analysis, both case and control samples must be handled according to the same protocol.

Temperature Considerations in Protein Sample Preparation

Temperature is a major variable in preparing protein samples. Protein degradation can be minimized by maintaining samples at low temperatures during processing and transport, meaning samples should be transported on dry ice to the Olink laboratory. The long-term storage temperature should be -80°C or lower.

Collection of Serum Samples

• Collect whole blood into serum collection tubes;
• Allow the blood to clot at room temperature, which typically takes 15-30 minutes. In samples left for over 60 minutes, the clotting cells may begin to dissolve;
• Centrifuge at 1000-2000 x g for 10 minutes in a refrigerated centrifuge (2-8°C) to remove the blood clot;
• Immediately transfer the serum to a clean tube;
• Store at -80°C.

Collection of Plasma Samples

• Collect whole blood into commercially available anticoagulant-treated collection tubes (treated with EDTA, citrate, or heparin);
• Centrifuge at 1000-2000 x g for 10 minutes in a refrigerated centrifuge (2-8°C) to remove cells from the plasma;
• Immediately transfer the plasma to a clean tube;
• Store at -80°C.

Types and Sizes of Collection Tubes

Any commercially available collection tubes specified for serum or plasma collection may be used. Ensure that all samples are collected using the same method and type of collection tube (same brand, same size, etc.).

3. Sample Transport

Shipping Tubes and Sizes

Sample should be sent in temperature-resistant, protein-non-binding plastic containers, such as 96-well PCR plates, preferably with a full skirt, and using a seal guaranteed to withstand -80°C. Olink laboratories can also transfer samples from PCR 8-tube strips or Eppendorf tubes to 96-well plates; please contact the laboratory in advance to confirm.

Sample Volume

Each sample should provide a minimum of 40 microliters (μL).

Recommended Volume

96-well plate: 50 microliters/sample

Eppendorf tube: 100-500 microliters/sample

Sample Quantity

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

Sample Labeling

Clearly label sample plates or tubes with temperature-resistant labels or permanent markers, for example, a simple alphanumeric code, so that they can be identified later (e.g., "A, B, C", "1, 2, 3", or "A1, A2, A3").

Transport Temperature

Samples should typically be transported on dry ice (i.e., they should still be frozen upon arrival).