Tau proteins and amyloid-β (Aβ) peptides are the current recognized cerebrospinal fluid (CSF) biomarkers used as an aid in the diagnosis of Alzheimer's disease (AD). However, there is no consensus on their clinical use due to non-qualified cut-off values, probably related to the observed high pre-analytical and analytical variability. Standardized pre-analytical protocols have therefore been proposed. Importantly, these recommend the use of polypropylene collection/sampling tubes while, to date, no broad comparison of these types of tubes has been conducted. In this study, we first compared, as part of a real clinical workflow, the impact of four different collection tubes on the CSF concentration of Aβ peptides (Aβ42, Aβ40) and total (hTau) and phosphorylated (P-Tau181P) tau proteins measured using routine ELISA kits. We then extended this study to 11 polypropylene tubes used by different clinical laboratories, and investigated their plastic polymer composition using differential scanning calorimetry and Fourier Transformed Infrared spectroscopy. Significant concentration variations linked solely to the use of different types of tubes were observed. This was particularly marked for Aβ peptides, with >50% disparity occurring in less than five minutes. Polymer composition analysis revealed that most polypropylene tubes were in fact copolymers with at least polyethylene. There was no clear correlation between tube composition and pre-analytical behavior. Our results show that the use of polypropylene tubes does not guarantee satisfactory pre-analytical behavior. They also point to collection/sampling tubes being a major pre-analytical source of variability that could impact the significance of AD biological diagnosis.
This review describes different strategies of surface elaboration for a better control of biomolecule adsorption. After a brief description of the fundamental interactions between surfaces and biomolecules, various routes of surface elaboration are presented dealing with the attachment of functional groups mostly thanks to plasma techniques, with the grafting to and from methods, and with the adsorption of surfactants. The grafting of stimuli-responsive polymers is also pointed out. Then, the discussion is focused on the protein adsorption phenomena showing how their interactions with solid surfaces are complex. The adsorption mechanism is proved to be dependent on the solid surface physicochemical properties as well as on the surface and conformation properties of the proteins. Different behaviors are also reported for complex multiple protein solutions.
The general properties of hydroxyethylcellulose (HEC) grafted with 3-glycidoxypropyltrimethoxysilane (GPTMS) or 3-glycidoxypropylmethyldiethoxysilane (GPDMS) were studied for potential biomedical applications. The graft involved a Williamson reaction between the free hydroxyl function of HEC and the epoxy function of the two silanes. As the grafted silanes are in ionic form (sodium silanolate), this product remains in gel form at basic pH (>12.3) in aqueous solution. When pH decreases, sodium silanolate is transformed into silanol (2 or 3 silanol functions are carried by silicon, depending on the silane grafted). The silanols interreact, and the gel is transformed into a cross-linking form at room or body temperature. Studies were conducted to optimise this product for specific uses. Steam sterilization was used to compare self-hardening as a function of the silane grafted. Our previous work indicated that HEC grafted with GPTMS has good reactivity, but requires high pH for dissolution, whereas dissolution occurs at lower pH with GPMDS. The rate of silanol condensation for silated HEC was then determined as a function of pH, temperature, type of silane, and the percentage grafted. Condensation rates were ascertained by the viscosity method, and gels were neutralized by different solutions to obtain buffered forms at various pH. The time required to obtain 10(5) mPa x s, with an initial state of 2500 mPa x s, was then calculated. Condensation was catalysed in acid or basic medium at a lower rate at pH 5.5-6.5, and a temperature rise increased the condensation rate, regardless of the pH or silane studied. Silanetriol was more reactive than silanediol. However, as HEC lost considerable viscosity after sterilization, further studies will be conducted to develop new polysaccharides grafted with silane.
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