Surface self-assembly of N-fluorenyl-9-methoxycarbonyl diphenylalanine on silica wafer

N-Fluorenyl-9-methoxycarbonyl diphenylalanine (Fmoc-FF-OH) was chemically immobilized to the surface of silica wafer as the "seed". When immersing this peptide attached silica wafer into the dipeptide aqueous solution, the occurrence of a pH triggered surface self-assembly resulted in the formation of peptide nanorods on the surface of silica wafer. This surface self-assembly exhibited a dependence on the concentration of the dipeptide aqueous solution. It was proposed that the self-assembly of this dipeptide on the surface of silica wafer was similar to that in aqueous solution. In comparison with the conventional physical adsorption on the substrates, the chemically attached self-assembled nanorods exhibited much improved adsorption capacity on the substrate surface.     

Biofunctionalization of a "clickable" organic layer photochemically grafted on titanium substrates

We have developed a general method combining photochemical grafting and copper-catalyzed click chemistry for biofunctionalization of titanium substrates. The UV-activated grafting of an α,ω-alkenyne onto TiO(2)/Ti substrates provided a "clickable" thin film platform. The selective attachment of the vinyl end of the molecule to the surface was achieved by masking the alkynyl end with a trimethylgermanyl (TMG) protecting group. Subsequently, various oligo(ethylene glycol) (OEG) derivatives terminated with an azido group were attached to the TMG-alkynyl modified titanium surface via a one-pot deprotection/click reaction. The films were characterized by X-ray photoelectron spectroscopy (XPS), contact angle goniometry, ellipsometry, and atomic force microscopy (AFM). We showed that the titanium surface presenting click-immobilized OEG substantially suppressed the nonspecific attachment of protein and cells as compared to the unmodified titanium substrate. Furthermore, glycine-arginine-glycine-aspartate (GRGD), a cell adhesion peptide, was coimmobilized with OEG on the platform. We demonstrated that the resultant GRGD-presenting thin film on Ti substrates can promote the specific adhesion and spreading of AsPC-1 cells.     

The activity of covalently immobilized Grubbs–Hoveyda type catalyst is highly dependent on the nature of the support material

A Hoveyda-type catalyst for olefin metathesis was synthesized and covalently attached via an amide bond to four different solid supports. One of these supports was a home-made hybrid silica support, where an ultra-thin copolymer of poly(styrene) and poly(acrylamide) was grafted on. The three other supports were commercially available, namely HypoGel 400, PEGA and Trisoperl. It was demonstrated that the catalysts were active in ring closing metathesis (RCM) reactions as well as in cross metathesis (CM) and ring opening metathesis (ROM) reactions, but the activity of the catalyst was highly dependent on the nature of the support.     

Physicochemical regularities of strontium sorption by sorbents based on di(<Emphasis Type="Italic">tert</Emphasis>-butyldicyclohexano)-18-crown-6

A number of sorbents based on di(tert-butyldicyclohexano)-18-crown-6, 1,1,7-trihydrododecafluoroheptanol, and various supports (Porolas-T, LPS-500, hydrophobized silica gel) were prepared. The effect of the supports on strontium sorption from nitric acid solutions was estimated. The physicochemical regularities of strontium sorption (isotherm, kinetics, and selectivity) were studied.     

Surface confined ionic liquid as a stationary phase for HPLC

Trimethoxysilane "ionosilane" derivatives of room temperature ionic liquids based on alkylimidazolium bromides were synthesized for attachment to silica support material. The derivatives 1-methyl-3-(trimethoxysilylpropyl)imidazolium bromide and 1-butyl-3-(trimethoxysilylpropyl)imidazolium bromide were used to modify the surface of 3 microm diameter silica particles to act as the stationary phase for HPLC. The modified particles were characterized by thermogravimetric analysis (TGA) and (13)C and (29)Si NMR spectroscopies. The surface modification procedure rendered particles with a surface coverage of 0.84 micromol m(-2) for the alkylimidazolium bromide. The ionic liquid moiety was predominantly attached to the silica surface through two siloxane bonds of the ionosilane derivative (63%). Columns packed with the modified silica material were tested under HPLC conditions. Preliminary evaluation of the stationary phase for HPLC was performed using aromatic carboxylic acids as model compounds. The separation mechanism appears to involve multiple interactions including ion exchange, hydrophobic interaction, and other electrostatic interactions.     

Rigid Support Materials for the Immobilization of Enzymes

The purpose of the work presented here was to prepare a support material for enzymes and “affinity ligands” with the following characteristics: low cost, durability, rigidity, and high capacity. Our study encompassed conjugates of porous and nonporous silicas with organic polymers and macroporous ion-exchange resins. Poly-ethyleneimine (PEI), polyacrylic acid (PAA), poly (methyl vinyl ether/maleic anhydride) were attached to porous glass and silica in various combinations. The composite of silica beads with PEI and PAA is a good support for the enzyme trypsin as judged by the activity against N-α-benzoyl-L-arginine ethyl ester.

Amberlyst (macroporous, sulfonated polystyrene) was activated by treatment with thionyl chloride; the resulting resin was either used directly or reacted with a diamine. The diamine derivative was used for enzyme coupling or transformed further to the succinyl or p-aminobenzoyl derivative. None of these derivatives were particularly good as supports for the enzyme trypsin. Duolite converted to a PAA, succinyl, or succinimide derivative was a good support. The enzyme-resin adduct has good activity and stability.

The resin is quite durable and of low cost. The Duolite-trypsin has good activity against protein. In addition, this derivative was active in 7 M urea. The proteolytic activity was nearly doubled by urea, presumably as a result of substrate (casein) denaturation. The michaelis constants and pH dependences are compared for trypsin conjugates with Duolite A-7, Silica-PEI-PAA, agarose, and porous glass. A cost comparison reveals that the Duolite and silica derivatives are much less expensive than agarose and glass.     

A "teardown" method to create large mesotunnels on the pore walls of ordered mesoporous silica

A "teardown" method to create large mesotunnels (approximately 9 nm) on the pore walls of ordered mesoporous silicas is demonstrated by digesting the organic constituents from polymer-silicate nanocomposites. The ordered mesostructured polymer-silicate composites were first obtained via the evaporation-induced triconstituent co-assembly method by using a low-molecular-weight phenolic resin (resols) as an organic precursor; prehydrolyzed TEOS as an inorganic precursor, and triblock copolymer F127 as a template. All of organic components including F127 and phenolic resins are removed by the microwave digestion (MWD) method from mesostructured polymer-silica composites. While the removal of triblock copolymer F127 generates main pore channels, the phenolic resins can also be torn down from the pore walls, yielding mesotunnels between the channels. The resulting silica products exhibit ordered 2-D hexagonal mesostructure, large pore volume (up to 1.92 cm(3)/g), and very large pore size (up to 22.9 nm), which is even larger than their mesostructural cell parameter (14.2 nm). TEM images confirm the existence of mesotunnels on the silica pore walls. FT-IR and (29)Si solid-state NMR results reveal that these silica products have a large number of silanol groups.     

Biofunctionalization on alkylated silicon substrate surfaces via "click" chemistry

Biofunctionalization of silicon substrates is important to the development of silicon-based biosensors and devices. Compared to conventional organosiloxane films on silicon oxide intermediate layers, organic monolayers directly bound to the nonoxidized silicon substrates via Si-C bonds enhance the sensitivity of detection and the stability against hydrolytic cleavage. Such monolayers presenting a high density of terminal alkynyl groups for bioconjugation via copper-catalyzed azide-alkyne 1,3-dipolar cycloaddition (CuAAC, a "click" reaction) were reported. However, yields of the CuAAC reactions on these monolayer platforms were low. Also, the nonspecific adsorption of proteins on the resultant surfaces remained a major obstacle for many potential biological applications. Herein, we report a new type of "clickable" monolayers grown by selective, photoactivated surface hydrosilylation of α,ω-alkenynes, where the alkynyl terminal is protected with a trimethylgermanyl (TMG) group, on hydrogen-terminated silicon substrates. The TMG groups on the film are readily removed in aqueous solutions in the presence of Cu(I). Significantly, the degermanylation and the subsequent CuAAC reaction with various azides could be combined into a single step in good yields. Thus, oligo(ethylene glycol) (OEG) with an azido tag was attached to the TMG-alkyne surfaces, leading to OEG-terminated surfaces that reduced the nonspecific adsorption of protein (fibrinogen) by >98%. The CuAAC reaction could be performed in microarray format to generate arrays of mannose and biotin with varied densities on the protein-resistant OEG background. We also demonstrated that the monolayer platform could be functionalized with mannose for highly specific capturing of living targets (Escherichia coli expressing fimbriae) onto the silicon substrates.     

Graft polymerization of styrene initiated by covalently bonded peroxide groups on silica

The graft polymerization of styrene initiated by immobilized peroxide groups was investigated. Three different types of modification reactions were used to introduce peroxide groups which are directly attached onto the surface of two different silica supports. Silanol groups were chlorinated using thionyl chloride or tetrachlorosilane. In another reaction pathway 1,3,5-benzenetricarbonyl chloride enabled the introduction of free acid chloride residues bonded onto the surface of silica. tert-Butyl hydroperoxide (TBHP) was used to transform the chlorosilyl and the acid chloride groups into peroxide residues. In a further reaction step the covalently bonded peroxides initiated the polymerization of styrene to form grafted polystyrene directly attached onto the silica support. Solid-state 13C CP/MAS NMR spectroscopy, and thermogravimetric and scanning electron microscope measurements enabled a clear structure and property elucidation of the different bonded phases. The highest amount of grafted polystyrene was achieved employing the acid chloride synthesis pathway with silica-gel, whereas modification of spherical silica only led to minor amounts of grafted polymer. The results contribute to the evolving need to understand particle surface modifications and may have positive impact on development of new HPLC stationary phases for improved elutant resolution.     

"Click" immobilization on alkylated silicon substrates: model for the study of surface bound antimicrobial peptides

We describe an effective approach for the covalent immobilization of antimicrobial peptides (AMPs) to bioinert substrates via Cu(I) -catalyzed azide-alkyne cycloaddition (CuAAC). The bioinert substrates were prepared by surface hydrosilylation of oligo(ethylene glycol) (OEG) terminated alkenes on hydrogen-terminated silicon surfaces. To render the OEG monolayers "clickable", mixed monolayers were prepared using OEG-alkenes with and without a terminal alkyne protected by a trimethylgermanyl (TMG) group. The mixed monolayers were characterized by X-ray photoelectron spectroscopy (XPS), elliposometry and contact angle measurement. The TMG protecting group can be readily removed to yield a free terminal alkyne by catalytic amounts of Cu(I) in an aqueous media. This step can then be combined with the subsequent CuAAC reaction. Thus, the immobilization of an azide modified AMP (N3-IG-25) was achieved in a one-pot deprotection/coupling reaction. Varying the ratio of the two alkenes in the deposition mixture allowed for control over the density of the alkynyl groups in the mixed monolayer, and subsequently the coverage of the AMPs on the monolayer. These samples allowed for study of the dependence of antimicrobial activities on the AMP density. The results show that a relative low coverage of AMPs (～1.6×10(13) molecule per cm(2)) is sufficient to significantly suppress the viability of Pseudomonas aeruginosa, while the surface presenting the highest density of AMPs (～2.8×10(13) molecule per cm(2)) is still cyto-compatible. The remarkable antibacterial activity is attributed to the long and flexible linker and the site-specific "click" immobilization, which may facilitate the covalently attached peptides to interact with and disrupt the bacterial membranes.     

Nickel catalysis for hydrogenation of <Emphasis Type="Italic">p</Emphasis>-dinitrobenzene to <Emphasis Type="Italic">p</Emphasis>-phenylenediamine

The activity of supported nickel catalysts (5–20% Ni) in the hydrogenation of p-dinitrobenzene to p-phenylenediamine was investigated. The catalysts were obtained by ureainduced precipitation. Activated carbon, alumina, titania, and silica gel were evaluated as supports. The most active catalysts, 5%Ni/TiO₂ and 20%Ni/SiO₂, provided 50–54% yields of p-phenylenediamine at complete dinitrobenzene conversion.     

"Click" dendrimers: synthesis, redox sensing of Pd(OAc)2, and remarkable catalytic hydrogenation activity of precise Pd nanoparticles stabilized by 1,2,3-triazole-containing dendrimers

"Click" dendrimers containing 1,2,3-triazolyl ligands that coordinate to PdII(OAc)2 have been synthesized in view of catalytic applications. Five of these dendrimers contain ferrocenyl termini directly attached to the triazole ligand in order to monitor the number of PdII that are introduced into the dendrimers by cyclic voltammetry. Reduction of the PdII-triazole dendrimers by using NaBH4 or methanol yields Pd nanoparticles (PdNPs) that are stabilized either by several dendrimers (G0, DSN) or by encapsulation inside a dendrimer (G1 and G2: DEN), as confirmed by TEM. Relative to PAMAM-DENs (PAMAM=poly(amidoamine)), the "click" DSNs and DENs show a remarkable efficiency and stability for olefin hydrogenation under ambient conditions of various substrates. The influence of the reductant of PdII bound to the dendrimers is dramatic, reduction with methanol leading to much higher catalytic activity than reduction with NaBH4. The most active NPs are shown to be those derived from dendrimer G1, and variation of its termini groups (ferrocenyl, alkyl, phenyl) allowed us to clearly delineate, optimize, and rationalize the role of the dendrimer frameworks on the catalytic efficiencies. Finally, hydrogenation of various substrates catalyzed by these PdNPs shows remarkable selectivity features.     

Covalent Attachment of Active Enzymes to Upconversion Phosphors Allows Ratiometric Detection of Substrates

Upconverting phosphors (UCPs) convert multiple low energy photons into higher energy emission via the process of photon upconversion and offer an attractive alternative to organic fluorophores for use as luminescent probes. Here, UCPs were capped with functionalized silica in order to provide a surface to covalently conjugate proteins with surface-accessible cysteines. Variants of green fluorescent protein (GFP) and the flavoenzyme pentaerythritol tetranitrate reductase (PETNR) were then attached via maleimide-thiol coupling in order to allow energy transfer from the UCP to the GFP or flavin cofactor of PETNR, respectively. PETNR retains its activity when coupled to the UCPs, which allows reversible detection of enzyme substrates via ratiometric sensing of the enzyme redox state.     

In flow activation of diol-silica with cyanogen bromide and triethylamine for preparing high-performance affinity chromatographic columns

A new coupling strategy using pre-packed diol-silica supports to obtain affinity columns for high-performance affinity chromatography (HPAC) is described. These columns were prepared by "in flow" activation in which solutions containing anhydrous solutions of CNBr and triethylamine are separately pumped to a mixer and then onto a pre-packed diol-silica column. Recycling the amino ligand to be coupled several times over the activated silica diol columns results in ligand immobilization. DNA (the Op 1 lac operator), 6-aminohexyl-Cibacron and a peptide (melittin) were all successfully "in flow" coupled to freshly activated columns. Methods for CNBr activation of pre-packed diol-silica column were developed for one, two or three pump HPLC systems. The supports were successfully used for the HPAC purification of a Lac repressor-beta-galactosidase fusion protein, alcohol dehydrogenase, and calmodulin. Columns prepared by in flow activation/coupling procedures were shown to be stable for at least 14 months. Also, in flow activated silica columns could be stored in anhydrous acetone for at least 3 months prior to coupling. Our experiments with these affinity ligand columns (DNA-silica, aminohexyl-Cibacron F3GA-silica, and melittin-silica), suggests that this is a very successful coupling protocol for producing a variety of HPAC columns.     

"Volatilizable" supports for high-throughput organic synthesis

The concepts and use of "volatilizable" solid supports are presented. Such supports, which are completely removed by volatilization following decomposition, improve the efficiency of the solid-phase synthesis of both individual and mixtures of low-molecular weight acyclic and heterocyclic compounds as well as peptides, peptidomimetics, and protected peptide fragments. The "volatilizable" silica-based solid supports and linkers used to illustrate this concept were found to be completely removed by their decomposition and ultimate "volatilization" during the final cleavage step of synthetic process to yield solely the desired synthetic product(s) in the final reaction vessel.     

OXIDATION OF HYDROCARBONS UNDER THE MILD CONDITIONS

The oxidation of organic substrates leads to the production of many functionalised molecules which are of great commercial and synthetic importance. The conventional mode of oxidation which involves stoichiometric ammount of Cr or Mn salts has been staked out because of the environmental hazadrous process. The transition to cleaner, safer, and more efficient plants is a new paradigm in the synthetic organic chemistry. Nowadays, hydrogen peroxide and oxygen as oxidizing agents were extreamely valuable and attractive. It is increasingly recognized, when polymers are used as supports for catalysts or organic reagents, the reactivity and selectivity of the supported catalysts or reagents may be seriously changed by so-called "polymer effects". As metal catalyzed oxidation of organic substrates with oxygen, we arc planing the incorporation of transition metals into polymer. In oxidaton organic compound has little resistant, so syntheses of organic-inorganic hybrid polymer from silica gel and montmorillonite by the modification with silane coupling reagents and the complexation of transition metal ions into hybrid polymer obtained above were investigated.     

Grafting of silica with a hydrophilic triol acrylamide polymer via surface-initiated "grafting from" method for hydrophilic-interaction chromatography

A novel hydrophilic polymer-coated silica sorbent has been prepared using the radical "grafting from" polymerization method through surface-bound azo initiators for hydrophilic-interaction chromatography (HILIC). The azo groups were introduced to the surface of silica gel through the reaction with amino groups on the surface of silica gel with 4,4'-azobis(4-cyanopentanoic acid chloride) (ACVC). The resultant azo-immobilized silica gel served as surface initiator to polymerize hydrophilic triol acrylamide monomer N-acryloyltris(hydroxymethyl) aminomethane (NA) in methanol to get hydrophilic polymer-coated silica sorbent. The obtained poly(NA)-coated silica (pNA-sil) was characterized by Fourier transform infrared spectroscopy (FT-IR), elemental analysis (EA), and nitrogen sorption porosimetry (NSP). Then the pNA-sil was packed into the stainless-steel column and evaluated in high-performance liquid chromatography (HPLC). Good chromatographic performance for the separation of peptides and nucleosides was obtained under HILIC mode. The results indicated that the pNA-sil stationary phase behaved as mixed-mode retention mechanisms of hydrophilic and ionic interactions. Furthermore, the pNA-sil phase was used to separate tryptic digest of β-casein and our results showed that more than 12 peptides peaks were resolved and well distributed within the elution window. Finally, the pNA-sil stationary phase was demonstrated to possess remarkable reproducibility and stability. Taken together, the pNA-sil stationary phase prepared in the current study offers a potential application in proteomics study.     

Immobilized penicillin G acylase as reactor and chiral selector in liquid chromatography

In this paper, the use of penicillin G acylase (PGA) as a biocatalyst and as a chiral selector is described. Penicillin G-acylase is an interesting enzyme used in the manufacture of semisynthetic antibiotics and, in particular, in the production of 6-APA by hydrolysis of penicillin G. Five PGA-based HPLC columns have been prepared by using two different silica supports by employing two immobilization methods, namely "in situ" and "in batch". The effects of the immobilization techniques and of different silica pore size on the catalytic properties of the enzyme as well as the applicability of the PGA-bonded stationary phases as chiral selectors for a number of chiral drugs have been investigated. The HPLC columns based on immobilized PGA combine the hydrolytic activity and the chiral recognition properties of PGA, therefore they have been used for the development of a combined reaction-separation system for chiral and achiral substrates.     

Surfaces Coated with Polymer Brushes Work as Carriers for Histidine Ammonia Lyase

The immobilization of enzymes on solid supports is an important challenge in biotechnology and biomedicine. In contrast to other methods, enzyme deposition in polymer brushes offers the benefit of high protein loading that preserves enzymatic activity in part due to the hydrated 3D environment that is available within the brush structure. The authors equipped planar and colloidal silica surfaces with poly(2-(diethylamino)ethyl methacrylate)-based brushes to immobilize Thermoplasma acidophilum histidine ammonia lyase, and analyzed the amount and activity of the immobilized enzyme. The poly(2-(diethylamino)ethyl methacrylate) brushes are attached to the solid silica supports either via a “grafting-to” or a “grafting-from” method. It is found that the grafting-from method results in higher amounts of deposited polymer and, consequently, higher amounts of Thermoplasma acidophilum histidine ammonia lyase. All polymer brush-modified surfaces show preserved catalytic activity of the deposited Thermoplasma acidophilum histidine ammonia lyase. However, immobilizing the enzyme in polymer brushes using the grafting-from method resulted in twice the enzymatic activity from the grafting-to approach, illustrating a successful enzyme deposition on a solid support.     

Direct printing of self-assembled lipid tubules on substrates

Lipid tubules formed by rolled-up bilayer sheets have shown promise in drug delivery systems, nanofluidics, and microelectronics. Here we report a method for directly printing lipid tubules on substrates. Preformed lipid tubules of 1,2-bis(tricosa-10,12-diynoyl)-sn-glycero-3-phosphocholine are aligned in the recessed channels of a thin poly(dimethylsiloxane) (PDMS) stamp. The aligned lipid tubules then serve as an "ink" for microcontact printing. We demonstrate that two-dimensional (2-D) arrays of aligned lipid tubules can be transferred onto planar, patterned, and curved substrates from the recessed channels of the PDMS stamp by bringing the tubule-inked PDMS stamp into contact with these substrates. We show that the 2-D array of aligned lipid tubules can be transcribed into a 2-D array of aligned silica cylinders through templated sol-gel condensation of tetraethoxysilane.