title: Correction of peripheral blood mononuclear cell cytosolic protein for hemoglobin contamination.
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.
authors: Division of Molecular Pathology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066CX Amsterdam, The Netherlands.
authors: Dick
authors: D
authors: Pluim
authors: Bart A W
authors: BA
authors: Jacobs
authors: Martin D
authors: MD
authors: Krähenbühl
authors: Anna E M
authors: AE
authors: Ruijter
authors: Jos H
authors: JH
authors: Beijnen
authors: Jan H M
authors: JH
authors: Schellens
mesh_terms: D003600:Cytosol / Q000737:chemistry* / Q000201:enzymology; D042943:Dihydrouracil Dehydrogenase (NADP) / Q000032:analysis; D005260:Female; D006454:Hemoglobins / Q000032:analysis*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000737:chemistry* / Q000201:enzymology; D008297:Male; D011506:Proteins / Q000032:analysis*; D013053:Spectrophotometry / Q000379:methods*
chemical_list: D006454:Hemoglobins; D011506:Proteins; D042943:Dihydrouracil Dehydrogenase (NADP)