Method validation for preparing serum and plasma samples from human blood for downstream proteomic, metabolomic, and...

Area of Science:

• Biotechnology
• Laboratory Science
• Biobanking

Background:

• Formal method validation for biospecimen processing is crucial for laboratory accreditation but often lacking.
• This study addresses the need for validated serum and plasma processing protocols.

Purpose of the Study:

• To validate serum and plasma processing protocols for fitness-for-purpose in downstream applications.
• To establish methodology for biospecimen processing method validation.

Main Methods:

• Optimized centrifugation conditions for serum and plasma preparation based on microparticle counts.
• Validated protocols for methodology and reproducibility using microparticle counts, DNA/hemoglobin concentration, and metabolomic/proteomic profiles.
• Evaluated robustness against variations in centrifugation temperature, deceleration, and pre-centrifugation delay.

Main Results:

• Optimal protocols: 10-min centrifugation for serum, 20-min for plasma at 2000 g, medium brake, room temperature.
• Protocols met criteria except for plasma metabolomics reproducibility.
• Room temperature processing yielded higher microparticles/DNA in serum and less hemolysis in plasma compared to 4°C.
• Protocols were robust for varying deceleration but sensitive to pre-centrifugation delay (2 hours) for microparticle counts.

Conclusions:

• Validated serum and plasma collection methods are suitable for protein, metabolite, and nucleic acid applications.
• Centrifugation temperature and pre-centrifugation delay significantly impact analytical results.
• Laboratories and biobanks must systematically record these processing conditions for accreditation.