Jansen Robert S, Rosing Hilde, Schellens Jan H M, Beijnen Jos H
Department of Pharmacy & Pharmacology, Slotervaart Hospital/The Netherlands Cancer Institute, Louwesweg 6, 1066, EC, Amsterdam, The Netherlands.
Anal Bioanal Chem. 2009 Oct;395(3):863-7. doi: 10.1007/s00216-009-3022-3. Epub 2009 Aug 16.
Quantitative analysis of intracellular analytes requires an accurate and precise assay not only for the quantitation of the analytes, but also for the quantitation of the number of cells in which they were determined. In this technical note we compare protein and DNA as markers for the number of peripheral blood mononuclear cells (PBMCs) isolated from whole blood. The protein content of samples was highly influenced by red blood cell contamination and was, therefore, a less suitable marker. The DNA-based method was unaffected by red blood cell contamination and was finally validated over a range from 10 x 10(6) to 300 x 10(6) PBMCs/mL.
细胞内分析物的定量分析不仅需要一种准确且精确的分析方法来定量分析物,还需要一种准确且精确的方法来定量测定分析物所在细胞的数量。在本技术说明中,我们比较了蛋白质和DNA作为从全血中分离的外周血单核细胞(PBMC)数量标志物的情况。样品的蛋白质含量受红细胞污染的影响很大,因此是一种不太合适的标志物。基于DNA的方法不受红细胞污染的影响,最终在10×10⁶至300×10⁶个PBMC/mL的范围内得到验证。