Identification and validation of platelet low biological variation proteins, superior to GAPDH, actin and tubulin, as tools in clinical proteomics.

摘要: Abstract Accurate biomarker quantification requires carefully chosen normalisation procedures. When single proteins are used as loading controls (LCs), it is crucial that their expressional stability must be known. Platelets an important source, especially for neurological diseases. We performed a systematical analysis of the platelet proteome to identify suitable LCs, using 2-D DIGE system. first screened healthy population (n = 137), aged between 18 and 104 years, find with small coefficients total variation (CV tot ), herein termed low biological (LBVP). Thereafter, was verified in 101 patients suffering from Alzheimer's- (AD), Parkinson's- disease, vascular dementia or schizophrenia. Interestingly, traditional LCs such tubulin beta-1 GAPDH, were not found amongst LBVP. The least variable protein, calculated over all 238 individuals, 14-3-3 gamma, CV 9.3%, showing no gender, age disease dependency. capability gamma superior LC quantifying Western blot signals AD-biomarker Monoamine Oxidase B patient versus controls. Similar results obtained HepG2 cells, treated vitro DNA-methyltransferase inhibitor 5-aza-2′deoxicytidine. Finally, we provide list alternative candidates accurate quantification. Biological significance This paper suggests considerable lower than well known “housekeeping” like GAPDH tubulin. Spot abundances middle ranged unaffected by certain Hence, listed might valuable additionally alternatively. Platelet's protein validated Furthermore demonstrated also stable expressed HepG2, cells others platelets, when DNA methylation inhibitor.