The protein corona (PC) that forms around nanomaterials upon exposure to humanbiofluids (e.g., serum, plasma, cerebral spinal fluid etc.) is personalized, i.e., it dependson alterations of the human proteome as those occurring in several cancer types.This may relevant for early cancer detection when changes in concentration of typicalbiomarkers are often too low to be detected by blood tests. Among nanomaterialsunder development for in vitro diagnostic (IVD) testing, Graphene Oxide (GO) isregarded as one of the most promising ones due to its intrinsic properties and peculiarbehavior in biological environments. While recent studies have explored the bindingof single proteins to GO nanoflakes, unexplored variables (e.g., GO lateral size andprotein concentration) leading to formation of GO-PC in human plasma (HP) haveonly marginally addressed so far. In this work, we studied the PC that forms aroundGO nanoflakes of different lateral sizes (100, 300, and 750 nm) upon exposure toHP at several dilution factors which extend over three orders of magnitude from 1(i.e., undiluted HP) to 103. HP was collected from 20 subjects, half of them beinghealthy donors and half of them diagnosed with pancreatic ductal adenocarcinoma(PDAC) a lethal malignancy with poor prognosis and very low 5-year survival rateafter diagnosis. By dynamic light scattering (DLS), electrophoretic light scattering (ELS),sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and nano liquidchromatography tandem mass spectrometry (nano-LC MS/MS) experiments we showthat the lateral size of GO has a minor impact, if any, on PC composition. On the otherside, protein concentration strongly affects PC of GO nanoflakes. In particular, we wereable to set dilution factor of HP in a way that maximizes the personalization of PC, i.e.,the alteration in the protein profile of GO nanoflakes between cancer vs. non-cancpatients. We believe that this study shall contribute to a deeper understanding of theinteractions among GO and HP, thus paving the way for the development of IVD toolsto be used at every step of the patient pathway, from prognosis, screening, diagnosis tomonitoring the progression of disease.
Personalized graphene oxide-protein corona in the human plasma of pancreatic cancer patients
Caputo D;Coppola R;
2020-01-01
Abstract
The protein corona (PC) that forms around nanomaterials upon exposure to humanbiofluids (e.g., serum, plasma, cerebral spinal fluid etc.) is personalized, i.e., it dependson alterations of the human proteome as those occurring in several cancer types.This may relevant for early cancer detection when changes in concentration of typicalbiomarkers are often too low to be detected by blood tests. Among nanomaterialsunder development for in vitro diagnostic (IVD) testing, Graphene Oxide (GO) isregarded as one of the most promising ones due to its intrinsic properties and peculiarbehavior in biological environments. While recent studies have explored the bindingof single proteins to GO nanoflakes, unexplored variables (e.g., GO lateral size andprotein concentration) leading to formation of GO-PC in human plasma (HP) haveonly marginally addressed so far. In this work, we studied the PC that forms aroundGO nanoflakes of different lateral sizes (100, 300, and 750 nm) upon exposure toHP at several dilution factors which extend over three orders of magnitude from 1(i.e., undiluted HP) to 103. HP was collected from 20 subjects, half of them beinghealthy donors and half of them diagnosed with pancreatic ductal adenocarcinoma(PDAC) a lethal malignancy with poor prognosis and very low 5-year survival rateafter diagnosis. By dynamic light scattering (DLS), electrophoretic light scattering (ELS),sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and nano liquidchromatography tandem mass spectrometry (nano-LC MS/MS) experiments we showthat the lateral size of GO has a minor impact, if any, on PC composition. On the otherside, protein concentration strongly affects PC of GO nanoflakes. In particular, we wereable to set dilution factor of HP in a way that maximizes the personalization of PC, i.e.,the alteration in the protein profile of GO nanoflakes between cancer vs. non-cancpatients. We believe that this study shall contribute to a deeper understanding of theinteractions among GO and HP, thus paving the way for the development of IVD toolsto be used at every step of the patient pathway, from prognosis, screening, diagnosis tomonitoring the progression of disease.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
