Home › Cell Biology › Preparation of Peripheral Blood Mononuclear Cell Pellets and Plasma from a Single Blood Draw at Clinical Trial Sites for Biomarker Analysis
Cell Biology JoVE (Open Access) Citable · DOI

Preparation of Peripheral Blood Mononuclear Cell Pellets and Plasma from a Single Blood Draw at Clinical Trial Sites for Biomarker Analysis

DOI: 10.3791/60776-v
What you'll learn
  • ✓ Prepare high-quality PBMC and plasma samples from single blood draw at clinical sites
  • ✓ Perform whole blood irradiation for DNA damage response biomarker analysis
  • ✓ Implement proper freezing and storage protocols for translational biomarker studies
Protocol

This protocol details clinically implementable preparation of high quality PBMC and plasma biosamples at the clinical trial site that can be used for translational biomarker analysis.

Difficulty
intermediate
Total time
~2-3 hours per blood draw (including irradiation, processing, and freezing steps)
Biosafety
BSL-1

Steps

1
Introduce PBMC and plasma collection methods

Overview of clinically implementable protocols for obtaining high-quality biosamples from single blood draw at trial sites for translational biomarker analysis.

▶ 00:05
2
Irradiate whole blood for DNA damage response

Expose whole blood samples to controlled irradiation to induce DNA damage response, enabling assessment of biomarker expression related to cellular stress responses.

▶ 01:08
3
Freeze PBMC pellets and plasma samples

Process and store prepared PBMC pellets and plasma at appropriate freezing conditions to preserve sample integrity for downstream biomarker analysis.

▶ 03:35
4
Perform Western blot analysis

Conduct Western blotting to detect and quantify biomarker proteins from processed PBMC samples.

▶ 04:45
5
Assess DNA damage response outcomes

Analyze representative results demonstrating DNA damage biomarker expression levels across treated and control samples.

▶ 05:43
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