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Updated: Apr 15, 2026
Author Spotlight: New Insights into PBMC Mitochondrial Responses Using Fluorespirometry
Published on: May 24, 2024
Immunocapture and microplate-based activity and quantity measurement of pyruvate dehydrogenase in human peripheral
Xiaowen Liu1, Hira Pervez, Lars W Andersen
1Department of Emergency Medicine, Beth Israel Deaconess Medical Center, One Deaconess Road, W/CC 2, Boston, MA 02215, USA.
Background:
Pyruvate dehydrogenase (PDH) activity is altered in many human disorders. Current methods require tissue samples and yield inconsistent results. We describe a modified method for measuring PDH activity from isolated human peripheral blood mononuclear cells (PBMCs). RESULTS/METHODOLOGY: We found that PDH activity and quantity can be successfully measured in human PBMCs. Freeze-thaw cycles cannot efficiently disrupt the mitochondrial membrane. Processing time of up to 20 h does not affect PDH activity with proteinase inhibitor addition and a detergent concentration of 3.3% showed maximum yield. Sample protein concentration is correlated to PDH activity and quantity in human PBMCs from healthy subjects.
Conclusion:
Measuring PDH activity from PBMCs is a novel, easy and less invasive way to further understand the role of PDH in human disease.
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