Electrospinning-based synthesis of highly ordered mesoporous silica fiber for lab-in-syringe enrichment of plasma peptides

Silica fiber with highly ordered mesoporous structure and continuously long fibrous property was synthesized on a large-scale for the first time. It can be applied to the rapid (less than 3 min) and effective enrichment of endogenous peptides with a novel lab-in-syringe approach.     

Gas chromatography of low molecular weight halocarbons on macroporous silica modified by a thin liquid phase film

Summary The use of the silica adsorbent Silochrom C-80 modified by a thin liquid stationary phase film is proposed for the gas chromatographic analysis of low molecular weight halocarbons. Such modified adsorbents are characterized by good retention of the volatile halocarbons and high selectivity with respect to halocarbons. The thermodynamic sorption characteristics of alkanes, chloroalkanes and chloromethanes on Silochrom modified by three liquid phases (Apiezon L, methylsilicone and polyethylene glycol 20M) and on Chromaton N with thick films of the same phases were studied. The heats of sorption (−ΔH) and free sorption energy changes (−ΔF) of these substances and the increments −Δ(ΔH) and −Δ(ΔF) per chlorine atom in the chloralkane molecule compared with n-alkanes with the same number of carbon atoms were determined. The “chlorine” retention indices of low molecular weight volatile halocarbons on three sorbents were determined to identify them in aqueous solutions using the electron-capture detector. The analysis of drinking water by this technique is demonstrated.     

Selective extraction of low molecular weight proteins by mesoporous silica particles with modified internal and external surfaces

A new mesoporous silica material with modified external and internal surfaces (alkyl diol–vinyl-SiO₂) was prepared and applied to selectively extract low molecular weight (LMW) proteins and peptides (less than 11 kDa) from biological samples. X-ray diffraction, N₂ adsorption, transmission electron microscopy, and Fourier transform infrared spectroscopy were used to characterize the alkyl diol–vinyl-SiO₂. This material was used to extract LMW proteins (peptides) from standard proteins, a mixture of peptides, and crude human plasma. We compared the amounts of different proteins adsorbed by alkyl diol–vinyl-SiO₂ and by vinyl-SiO₂ (vinyl group on the internal surface of the mesoporous silica material). The desorption recovery of alkyl diol–vinyl-SiO₂ by different eluents was also investigated by using a standard protein (insulin) as a model LMW protein. The material could efficiently extract LMW proteins and peptides from a mixture of standard proteins and crude human plasma with good extraction efficiency and desorption recovery. Furthermore, the alkyl diol–vinyl-SiO₂ had much better extraction selectivity for LMW proteins and peptides than a commercial C₁₈ solid phase extraction material.     

Manipulation of molecular transport into mesoporous silica thin films by the infiltration of polyelectrolytes

The design of hybrid mesoporous materials incorporating polymeric assemblies as versatile functional units has become a very fertile research area offering major opportunities for controlling molecular transport through interfaces. However, the creation of such functional materials depends critically on our ability to assemble polymeric units in a predictable manner within mesopores with dimensions comparable to the size of the macromolecular blocks themselves. In this work, we describe for the first time the manipulation of the molecular transport properties of mesoporous silica thin films by the direct infiltration of polyelectrolytes into the inner environment of the 3D porous framework. The hybrid architectures were built up through the infiltration-electrostatic assembly of polyallylamine (PAH) on the mesopore silica walls, and the resulting systems were studied by a combination of experimental techniques including ellipso-porosimetry, cyclic voltammetry and X-ray photoelectron spectroscopy, among others. Our results show that the infiltration-assembly of PAH alters the intrinsic cation-permselective properties of mesoporous silica films, rendering them ion-permeable mesochannels and enabling the unrestricted diffusion of cationic and anionic species through the hybrid interfacial architecture. Contrary to what happens during the electrostatic assembly of PAH on planar silica films (quantitative charge reversal), the surface charge of the mesoporous walls is completely neutralized upon assembling the cationic PAH layer (i.e., no charge reversal occurs). We consider this work to have profound implications not only on the molecular design of multifunctional mesoporous thin films but also on understanding the predominant role of nanoconfinement effects in dictating the functional properties of polymer-inorganic hybrid nanomaterials.     

Comparison of three different enrichment strategies for serum low molecular weight protein identification using shotgun proteomics approach

Serum low-molecular weight (LMW) proteins potentially contain useful biological information and their identification can be used to discover novel potential biomarkers. Given the high complexity of serum samples, in the last years several different prefractionation and enrichment strategies have been developed. In this study three different methods, i.e. hydrogel nanoparticles, Proteominer^® peptide ligand affinity beads and Sartorius Vivaspin^® centrifugal ultrafiltration device, were compared and evaluated in order to select the best strategy for the enrichment and prefractionation of LMW proteins. A shotgun proteomics approach was adopted, with in-solution proteolytic digestion of the whole protein mixture and determination of the resulting peptides by nanoHPLC coupled with a high-resolution Orbitrap LTQ-XL mass spectrometer. Data analysis, focusing on the LMW proteome (MW ≤ 40 kDa), has shown that the hydrogel nanoparticles performed better in enriching the LMW protein profiles, with 115 proteins identified against 93 and 95 for Proteominer^® beads and Sartorius Vivaspin^® device, respectively.     

Tuning of Ti-doped mesoporous silica for highly efficient enrichment of phosphopeptides in human placenta mitochondria

Extraction of phosphopeptides from rather complex biological samples has been a tough issue for deep and comprehensive investigation into phosphoproteomes. In this paper, we present a series of Ti-doped mesoporous silica (Ti-MPS) materials with tunable composition and controllable morphology for highly efficient enrichment of phosphopeptides. By altering the molar ratio of silicon to titanium (Si/Ti) in the precursor, the external morphology, Ti content, internal long-rang order, and surface area of Ti-MPS were all modulated accordingly with certain regularity. Tryptic digests of standard phosphoprotein α- and β-casein were employed to assess the phosphopeptide enrichment capability of Ti-MPS series. At the Si/Ti molar ratio of 8:1, the optimum enrichment performance with admirable sensitivity and capacity was achieved. The detection limit for β-casein could reach 10 fmol, and 15 phosphopeptides from the digest of α-casein were resolved in the spectrum after enrichment, both superior to the behavior of commercial TiO₂ materials. More significantly, for the digest of human placenta mitochondria, 396 phosphopeptides and 298 phosphoproteins were definitely detected and identified after enrichment with optimized Ti-MPS material, demonstrating its remarkable applicability for untouched phosphoproteomes. In addition, this research also opened up a universal pathway to construct a composition-tunable functional material in pursuit of the maximum performance in applications.

Synthesis of cobalt doped silica thin film for low temperature optical gas sensor

A modified sol–gel method was used to prepare cobalt doped silica thin film with a cobalt content of 10, 20 and 30 mol% (10Co, 20Co and 30Co). The prepared films were annealed at different temperatures in the range 400–1,000 °C, and their structural evolution examined. The mixed valence cobalt oxide, Co₃O₄, crystallizes only in the sample with the higher cobalt content, while cobalt silicate is the only crystalline phase detected in the sample 10Co and 20Co. Both the cobalt content and the temperature of heat treatment resulted to affect the nature of cobalt species dispersed in the silica matrix. The 30Co was selected for further investigations by FTIR spectroscopy to follow the structural evolution of 30Co film as function of the temperature and UV–Vis to get information on the cobalt valence state. The optical gas-sensing properties of 30Co films, containing Co₃O₄ as the major cobalt phase, were studied through the measuring of the film transmittance in dry air and in presence of dry air containing variable concentrations of polluting gases, CO and NO₂. The 30Co samples resulted to be highly sensitive to CO at room temperature. An explanation for the CO sensing characteristics, at low temperature, was proposed by referring to the physisorption-related mechanics of CO.     

Unmasking low-abundance peptides from human blood plasma and serum samples by a simple and robust two-step precipitation/immunoaffinity enrichment method

Proteomic analysis of human plasma and serum for identifying and validating disease-specific marker proteins and peptides has one major drawback besides its unique advantage as a readily available sample source for diagnostic assays. This disadvantage is represented by the predominance of several high- and middle-abundant proteins, which clearly hamper identification and quantification approaches of potential and validated protein and peptide biomarkers, which are often of very low abundance. During the last decades, a significant number of depletion and enrichment techniques evolved to address these two issues. We present here a cost-effective and easy-to-use strategy for protein depletion comprising a thermal precipitation protocol followed by a two-step liquid/liquid precipitation as well as using an immunoaffinity chromatography method for the specific enrichment and isolation of the low-abundance polypeptide N-terminal pro-B-type natriuretic peptide and its precursor proBNP clinically used as biomarkers for the detection of severe human heart failure and related diseases. The applicability of this approach is shown by SDS -CGE, SDS-PAGE, electrochemiluminescence immunoassay and nano-LC ESI-MS/MS. Our thermal precipitation protocol followed by a two-step liquid/liquid precipitation could also serve as a potential depletion technique for the characterization of other low-abundance peptides and proteins.     

Improved selection of LMW over HMW proteins from human plasma by mesoporous silica particles with external modification

Selective extraction of low molecular weight (LMW) proteins and peptides from complex biological samples plays an important role in the discovery of useful biomarkers and signaling molecules. Various methods, such as solid-phase extraction (SPE), ultrafiltration, and size-exclusion chromatography have been developed for such extraction purpose. In this study, we present, to our knowledge, the first demonstration of alkyl-diol@SiO₂ mesoporous material MCM-41 (alkyl-diol group on the external surface of mesoporous material) for selective extraction of LMW proteins and peptides from complex biological samples. The adsorption kinetics of LMW proteins, the influence of pH on adsorption and the desorption recovery by different elution solvents were investigated by using standard proteins as model samples. It was demonstrated that the modification of alkyl-diol group on the external surface could efficiently decrease the adsorption of HMW protein and increase the desorption recovery of LMW protein. Furthermore, the mesoporous materials were applied to selectively extract LMW proteins and peptides (<10 kDa) from crude human plasma. And the modified MCM-41 material had much better extraction selectivity and efficiency for LMW proteins and peptides than unmodified one.     

Improved disc SDS‐PAGE for extraction of low molecular weight proteins from serum

The low molecular weight proteins can provide a lot of valuable information of biomarkers. To study these proteins, the high abundance and high molecular weight proteins must be removed prior to analysis. In this work, a simple and inexpensive disc SDS‐PAGE to extract low molecular weight proteins from human serum and cutoff proteins larger than 30 kDa was developed. Some experimental conditions were examined. The experimental results obtained by plate SDS‐PAGE and MALDI‐TOF MS showed that the molecular weight of extracted proteins was about in the range from 0.3 to 28 kDa. Some experiments, including precipitation of proteins in organic solvents, SPE and cytochrome C test, were carried out and the experimental results demonstrated successful recovery of proteins/peptides with molecular weight from several hundreds of dalton to about 30 kDa. The experimental results obtained by plate SDS‐PAGE indicated the repeatability was satisfactory.     

Preparation of nano-sized Pt metal particles by photo-assisted deposition (PAD) on transparent Ti-containing mesoporous silica thin film

We have investigated a novel technique for the preparation of nano-sized Pt metals on Ti-containing mesoporous silica (TMS) thin film by photo-assisted deposition (PAD). The transparent TMS thin film was prepared on a quartz plate through sol—gel/spin coating. XRD, UV-Vis and Ti K-edge XAFS measurements revealed the formation of isolated Ti oxide species with a tetrahedral-coordination geometry in the silica framework. Deposition of Pt metal precursor on TMS thin film under UV-light irradiation, followed by reduction with molecular hydrogen, afforded a transparent thin film (Pt/TMS). The formation of highly dispersed nano-sized Pt metals having narrow size distributions was determined by Pd L_III-edge XANES and TEM analysis. The TMS and Pt/TMS thin films have been demonstrated to exhibit a strong hydrophilic property, even before UV irradiation, compared to the common mesoporous silica and TiO₂ thin films. After UV-light irradiation, the contact angle of water droplet on the TMS and Pt/TMS thin films became extremely lower, indicating the appearance of the photo-induced super-hydrophilic property.     

Highly transparent superhydrophobic thin film with low refractive index prepared by one-step coating of modified silica nanoparticles

An easy and effective method to prepare superhydrophobic thin film has been developed. The film with optically transparent and low refractive index was composed by one-step coating with modified silica nanoparticles. The silica nanoparticles were prepared by sol–gel process of hydrolysis and condensation of alkoxysilane compounds and then surface modification silica nanoparticles, 50 ± 10 nm, were accomplished using methoxytrimethylsilane (MOTMS). Water contact angle of film increased with the weight of MOTMS of silica sol. When the weight of MOTMS was optimized, the water contact angle and sliding angle of film were 152.8° and less than 10°, respectively. The transmittance of film was also increased as compared to the un-coated microscope glass slide, from 91 to 93.5 %. The refractive index of the film was approximately 1.09 as measured by ellipsometer. The superhydrphobic thin film was also successfully made by using spray coating and the water contact angle of this film was more than 160°. Surface morphology of difference coating methods, dip and spray, were studied. Our result suggests that the film can be applied for superhydrophobicity and optical applications.     

Comparison of three methods for fractionation and enrichment of low molecular weight proteins for SELDI-TOF-MS differential analysis

In most diseases, the clinical need for serum/plasma markers has never been so crucial, not only for diagnosis, but also for the selection of the most efficient therapies, as well as exclusion of ineffective or toxic treatment. Due to the high sample complexity, prefractionation is essential for exploring the deep proteome and finding specific markers.In this study, three different sample preparation methods (i.e., highly abundant protein precipitation, restricted access materials (RAM) combined with IMAC chromatography and peptide ligand affinity beads) were investigated in order to select the best fractionation step for further differential proteomic experiments focusing on the LMW proteome (MW inferior to 40,000 Da). Indeed, the aim was not to cover the entire plasma/serum proteome, but to enrich potentially interesting tissue leakage proteins. These three methods were evaluated on their reproducibility, on the SELDI-TOF-MS peptide/protein peaks generated after fractionation and on the information supplied.The studied methods appeared to give complementary information and presented good reproducibility (below 20%). Peptide ligand affinity beads were found to provide efficient depletion of HMW proteins and peak enrichment in protein/peptide profiles.     

Performance evaluation of functionalized ordered mesoporous silica for VOCs adsorption by molecular dynamics simulation

Volatile organic compounds (VOCs) are growing pollutants now that cause the serious environmental pollution and threaten human health. The functionalized ordered mesoporous silica (FOMS) has attracted considerable attention in adsorbing VOCs. In this paper, the molecular dynamics simulation was used to simulate the adsorption performance of FOMS on VOCs (acetone, ethyl acetate and toluene). After simulating different pore sizes (2 nm, 3 nm and 4 nm) adsorption performances of ordered mesoporous silica (OMS) on VOCs, OMS with a pore size of 4 nm was selected to further study the influence of functional groups (vinyl, methyl, and phenyl). The following law was obtained: the saturated adsorption capacities of vinyl-functionalized OMS (V-FOMS) to acetone, ethyl acetate and toluene were 3.045 mmol.g^?1, 2.568 mmol.g^?1 and 1.976 mmol.g^?1 respectively; the saturated adsorption capacities of methyl-functionalized OMS (M-FOMS) to acetone, ethyl acetate and toluene were 2.798 mmol.g^?1, 2.312 mmol.g^?1 and 1.698 mmol.g^?1 respectively; the saturated adsorption capacities of phenyl-functionalized OMS (P-FOMS) to acetone, ethyl acetate and toluene were 2.124 mmol.g^?1, 1.941 mmol.g^?1 and 1.539 mmol.g^?1 respectively. These results show that the adsorption ability of FOMS for different adsorbates follows the sequence of acetone > ethyl acetate > toluene. Furthermore, the interaction between functional groups (vinyl, methyl and phenyl) in FOMS and VOCs was explored. It is found that the interaction between different functional groups and adsorbates is different (interaction energy effect). This interaction energy effect promotes FOMS to better adsorb VOCs. This work would provide fundamental understanding and guidance for the development of novel adsorption materials for the adsorption of VOCs.     

Preparation of phenyl‐functionalized magnetic mesoporous silica microspheres for the fast separation and selective enrichment of phenyl‐containing peptides

Peptide enrichment before mass spectrometry analysis is essential for large‐scale peptidomic studies, but challenges still remain. Herein, magnetic mesoporous silica microspheres with phenyl group modified interior pore walls were prepared by a facile sol–gel coating strategy, and were successfully applied for selective enrichment of phenyl‐containing peptides in complex biological samples. The newly prepared nanomaterials possessed abundant silanol groups in the exterior surface and numerous phenyl groups in the interior pore walls, as well as a large surface area (592.6 m²/g), large pore volume (0.33 cm³/g), uniform mesopores (3.8 nm), strong magnetic response (29.3 emu/g), and good dispersibility in aqueous solution. As a result of the unique structural properties and size‐exclusion effect, the core–shell phenyl‐functionalized magnetic mesoporous silica microspheres exhibited excellent performance in fast separation and selective enrichment of phenyl‐containing peptides, and the adsorption capacity for bradykinin reached 22.55 mg/g. In addition, selective enrichment of phenyl‐containing peptides from complex samples that are consist of peptides, large proteins, and human serum were achieved by using the as‐prepared microspheres, followed by high‐performance liquid chromatography with ultraviolet detection and electrospray ionization quadrupole time‐of‐flight mass spectrometry analysis. These results demonstrated the as‐prepared microspheres would be a potential candidate for endogenous phenyl‐containing peptides enrichment and biomarkers discovery in peptidome analysis.     

Molecular simulation of model sulfated polysaccharides of low molecular weight from Ganoderma lucidum and their interaction with human serum albumin

The extracellular polysaccharides from Ganoderma lucidum possess low molecular weight and are mainly composed of (1 → 3)/(1 → 6) linked-α-glucan and (1 → 3)/(1 → 6) linked-α-galactose. In the present study, conformations of the polysaccharides were simulated and investigated using Discovery studio 2.5. Their sulfated derivatives were also taken into account. The simulation was performed using CHARMM force field. The results show that the polysaccharides exist as random coil conformations, while the sulfated derivatives adopt more extensive and stiffer conformations in most cases. In addition, the molecular docking between polysaccharide and human serum albumin (HSA) was also investigated using ZDOCK module in Discovery studio 2.5. It is shown that the unmodified polysaccharide possesses higher affinity with HSA than the sulfated derivatives do due to its higher ZDOCK score.