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Correction of peripheral blood mononuclear cell cytosolic protein for hemoglobin contamination

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Abstract

Pharmacodynamic (PD) analysis requires accurate and precise quantification of enzyme activity targeted by anticancer agents in surrogate cells like peripheral blood mononuclear cells (PBMCs). Enzyme activity is normally reported per mass unit of protein input. However, high and fluctuating hemoglobin (Hb) contamination strongly influences the protein content of PBMC cytosolic lysate. We present the development and validation of a spectrophotometrical Hb quantification method to correct for this contamination. The applicability of Hb correction was demonstrated by determination of the dihydropyrimidine dehydrogenase enzyme activity in PBMC cytosolic lysates.

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Authors and Affiliations

  1. Division of Molecular Pathology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066CX, Amsterdam, The Netherlands

    Dick Pluim, Bart A. W. Jacobs, Martin D. Krähenbühl, Anna E. M. Ruijter & Jan H. M. Schellens

  2. Department of Pharmacy and Pharmacology, Slotervaart Hospital/The Netherlands Cancer Institute, Louwesweg 6, 1066EC, Amsterdam, The Netherlands

    Jos H. Beijnen

  3. Division Pharmacoepidemiology and Clinical Pharmacology, Department of Pharmaceutical Sciences, Science Faculty, University of Utrecht, Sorbonnelaan 14-16, 3584CA, Utrecht, The Netherlands

    Jos H. Beijnen & Jan H. M. Schellens

Authors
  1. Dick Pluim
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  2. Bart A. W. Jacobs
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  3. Martin D. Krähenbühl
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  4. Anna E. M. Ruijter
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  5. Jos H. Beijnen
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  6. Jan H. M. Schellens
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Corresponding author

Correspondence to Jan H. M. Schellens.

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Pluim, D., Jacobs, B.A.W., Krähenbühl, M.D. et al. Correction of peripheral blood mononuclear cell cytosolic protein for hemoglobin contamination. Anal Bioanal Chem 405, 2391–2395 (2013). https://doi.org/10.1007/s00216-012-6614-2

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  • DOI: https://doi.org/10.1007/s00216-012-6614-2

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