• Bioanalysis · Jan 2015

    Immunocapture and microplate-based activity and quantity measurement of pyruvate dehydrogenase in human peripheral blood mononuclear cells.

    • Xiaowen Liu, Hira Pervez, Lars W Andersen, Amy Uber, Sophia Montissol, Parth Patel, and Michael W Donnino.
    • Department of Emergency Medicine, Beth Israel Deaconess Medical Center, One Deaconess Road, W/CC 2, Boston, MA 02215, USA.
    • Bioanalysis. 2015 Jan 1; 7 (5): 583-92.

    BackgroundPyruvate 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.ConclusionMeasuring 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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