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Protein Delivery (protein + delivery)
Selected AbstractsSelf-Assembled Nanogel of Hydrophobized Dendritic Dextrin for Protein DeliveryMACROMOLECULAR BIOSCIENCE, Issue 7 2009Yayoi Ozawa Abstract Highly branched cyclic dextrin derivatives (CH-CDex) that are partly substituted with cholesterol groups have been synthesized. The CH-CDex forms monodisperse and stable nanogels with a hydrodynamic radii of ,10 nm by the self-assembly of 4,6 CH-CDex macromolecules in water. The CH-CDex nanogels spontaneously trap 10,16 molecules of fluorescein isothiocyanate-labeled insulin (FITC-Ins). The complex shows high colloidal stability: no dissociation of trapped insulin is observed after at least 1 month in phosphate buffer (0.1 M, pH 8.0). In the presence of bovine serum albumin (BSA, 50 mg,·,mL,1), which is a model blood system, the FITC-Ins trapped in the nanogels is continuously released (,20% at 12 h) without burst release. The high-density nanogel structure derived from the highly branched CDex significantly affects the stability of the nanogel,protein complex. [source] Examination of intravenous and intra-CSF protein delivery for treatment of neurological diseaseEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 6 2009Kim M. Hemsley Abstract Mucopolysaccharidosis type IIIA is a neurodegenerative lysosomal storage disorder characterized by progressive loss of learned skills, sleep disturbance and behavioural problems. Absent or greatly reduced activity of sulphamidase, a lysosomal protein, results in intracellular accumulation of heparan sulphate. Subsequent neuroinflammation and neurodegeneration typify this and many other lysosomal storage disorders. We propose that intra-cerebrospinal fluid protein delivery represents a potential therapeutic avenue for treatment of this and other neurodegenerative conditions; however, technical restraints restrict examination of its use prior to adulthood in mice. We have used a naturally-occurring Mucopolysaccharidosis type IIIA mouse model to determine the effectiveness of combining intravenous protein replacement (1 mg/kg) from birth to 6 weeks of age with intra-cerebrospinal fluid sulphamidase delivery (100 ,g, fortnightly from 6 weeks) on behaviour, the level of heparan sulphate-oligosaccharide storage and other neuropathology. Mice receiving combination treatment exhibited similar clinical improvement and reduction in heparan sulphate storage to those only receiving intra-cerebrospinal fluid enzyme. Reductions in micro- and astrogliosis and delayed development of ubiquitin-positive lesions were seen in both groups. A third group of intravenous-only treated mice did not exhibit clinical or neuropathological improvements. Intra-cerebrospinal fluid injection of sulphamidase effectively, but dose-dependently, treats neurological pathology in Mucopolysaccharidosis type IIIA, even when treatment begins in mice with established disease. [source] Type III secretion: The bacteria-eukaryotic cell expressFEMS MICROBIOLOGY LETTERS, Issue 1 2005Luís Jaime Mota Abstract Type III secretion (T3S) is an export pathway used by Gram-negative pathogenic bacteria to inject bacterial proteins into the cytosol of eukaryotic host cells. This pathway is characterized by (i) a secretion nanomachine related to the bacterial flagellum, but usually topped by a stiff needle-like structure; (ii) the assembly in the eukaryotic cell membrane of a translocation pore formed by T3S substrates; (iii) a non-cleavable N-terminal secretion signal; (iv) T3S chaperones, assisting the secretion of some substrates; (v) a control mechanism ensuring protein delivery at the right place and time. Here, we review these different aspects focusing in open questions that promise exciting findings in the near future. [source] Protein-Release Behavior of Self-Assembled PEG,, -Cyclodextrin/PEG,Cholesterol HydrogelsADVANCED FUNCTIONAL MATERIALS, Issue 18 2009Frank van de Manakker Abstract This paper reports on the degradation and protein release behavior of a self-assembled hydrogel system composed of , -cyclodextrin- (,CD) and cholesterol-derivatized 8-arm star-shaped poly(ethylene glycol) (PEG8). By mixing ,CD- and cholesterol-derivatized PEG8 (molecular weights 10, 20 and 40 kDa) in aqueous solution, hydrogels with different rheological properties are formed. It is shown that hydrogel degradation is mainly the result of surface erosion, which depends on the network swelling stresses and initial crosslink density of the gels. This degradation mechanism, which is hardly observed for other water-absorbing polymer networks, leads to a quantitative and nearly zero-order release of entrapped proteins. This system therefore offers great potential for protein delivery. [source] Hydrogels in Regenerative Medicine,ADVANCED MATERIALS, Issue 32-33 2009Brandon V. Slaughter Abstract Hydrogels, due to their unique biocompatibility, flexible methods of synthesis, range of constituents, and desirable physical characteristics, have been the material of choice for many applications in regenerative medicine. They can serve as scaffolds that provide structural integrity to tissue constructs, control drug and protein delivery to tissues and cultures, and serve as adhesives or barriers between tissue and material surfaces. In this work, the properties of hydrogels that are important for tissue engineering applications and the inherent material design constraints and challenges are discussed. Recent research involving several different hydrogels polymerized from a variety of synthetic and natural monomers using typical and novel synthetic methods are highlighted. Finally, special attention is given to the microfabrication techniques that are currently resulting in important advances in the field. [source] PTD4-apoptin protein therapy inhibits tumor growth in vivoINTERNATIONAL JOURNAL OF CANCER, Issue 12 2009Jun Sun Abstract Apoptin protein harbors tumor-selective cell death activity, which makes it a potential anticancer therapy candidate. This study reports an apoptin therapy approach based on protein transduction domain 4 (PTD4)-mediated transduction of recombinant apoptin protein. In vitro, the PTD4-apoptin fusion protein is located in the nucleus and induces cell death in, e.g., human hepatocarcinoma HepG2 cells. In normal human L-02 hepatocytes, PTD4-apoptin protein retained mainly cytoplasmic and did not induce detectable levels of cell death, illustrating that the PTD4 domain does not affect apoptin's tumor-selective characteristics. In vivo, liver, cervix and gastric carcinoma xenografts treated with PTD4-apoptin protein for 6 days via the tumor epidermis exhibited a significant tumor growth inhibition because of apoptin-mediated cell death. In addition, treatment of human hepatocarcinoma xenografts during 3 weeks showed that PTD4-apoptin protein has significant anticancer activity, whereas control treatment with PTD4-enhanced green fluorescence protein or saline did not. Cell death and disruption of the tumor integrity were apparent in the PTD4-apoptin transduced xenografted tumors. As important, although PTD4-apoptin protein could be detected in the epidermal tissue covering the subcutaneous tumor tissue and in several organs, such as liver and brain, of the treated mice, no tissue disruption or signs of cell death could be detected. Our in vivo data reveal that apoptin protein delivery constitutes a novel powerful and safe anticancer therapy. © 2009 UICC [source] Split target specificity of ResT: a design for protein delivery, site selectivity and regulation of enzyme activity?MOLECULAR MICROBIOLOGY, Issue 3 2007Makkuni Jayaram Summary The ResT telomere resolvase is responsible for maintaining the hairpin telomeres that cap the linear chromosome and minichromosomes of Borrelia burgdorferi. This enzyme acts at the tandem telomere junctions present within circular dimers resulting from DNA replication. ResT mediates the transesterification steps of resolution using a constellation of active site residues similar to that found in tyrosine recombinases and type IB topoisomerases. By combining this reaction mechanism with a hairpin binding module in its N-terminal domain, ResT reduces a fused telomere dimer into two hairpin monomers. ResT displays a split DNA binding specificity, with the N- and C-terminal domains targeting distinct regions of the telomere. This bi-specificity in binding is likely to be important in protein delivery, substrate selection and regulation of enzyme activity. [source] Single walled carbon nanotubes for transport and delivery of biological cargosPHYSICA STATUS SOLIDI (B) BASIC SOLID STATE PHYSICS, Issue 13 2006Nadine Wong Shi Kam Abstract The innate ability of carbon nanotubes to breach the cell membrane of various types of mammalian cells has been reported. Here, we present a summary of the various applications of carbon nanotubes as a cellular transport and delivery system for functional biological cargos. The internalization of SWNTs for transport and delivery into cells is mediated via endocytosis and does not appear to have any detrimental effect on either the transported cargo or the breached cell. The emergence of SWNT as a new class of cellular transporters holds many exciting promises for SWNT-based systems for drug delivery, protein delivery, gene therapy and cancer therapy applications. (© 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source] Networks for recognition of biomolecules: molecular imprinting and micropatterning poly(ethylene glycol)- Containing films,POLYMERS FOR ADVANCED TECHNOLOGIES, Issue 10-12 2002Mark E. Byrne Abstract Engineering the molecular design of biomaterials by controlling recognition and specificity is the first step in coordinating and duplicating complex biological and physiological processes. Studies of protein binding domains reveal molecular architectures with specific chemical moieties that provide a framework for selective recognition of target biomolecules in aqueous environment. By matching functionality and positioning of chemical residues, we have been successful in designing biomimetic polymer networks that specifically bind biomolecules in aqueous environments. Our work addresses the preparation, behavior, and dynamics of the three-dimensional structure of biomimetic polymers for selective recognition via non-covalent complexation. In particular, the synthesis and characterization of recognitive gels for the macromolecular recognition of D -glucose is highlighted. Novel copolymer networks containing poly(ethylene glycol) (PEG) and functional monomers such as acrylic acid, 2-hydroxyethyl methacrylate, and acrylamide were synthesized in dimethyl sulfoxide (polar, aprotic solvent) and water (polar, protic solvent) via UV-free radical polymerization. Polymers were characterized by single and competitive equilibrium and kinetic binding studies, single and competitive fluorescent and confocal microscopy studies, dynamic network swelling studies, and ATR-FTIR. Results qualitatively and quantitatively demonstrate effective glucose-binding polymers in aqueous solvent. Owing to the presence of template, the imprinting process resulted in a more macroporous structure as exhibited by dynamic swelling experiments and confocal microscopy. Polymerization kinetic studies suggest that the template molecule has more than a dilution effect on the polymerization, and the effect of the template is related strongly to the rate of propagation. In addition, PEG containing networks were micropatterned to fabricate microstructures, which would be the basis for micro-diagnostic and tissue engineering devices. Utilizing photolithography techniques, polymer micropatterns of a variety of shapes and dimensions have been created on polymer and silicon substrates using UV free-radical polymerizations with strict spatial control. Micropatterns were characterized using optical microscopy, SEM, and profilometry. The processes and analytical techniques presented are applicable to other stimuli-sensitive and recognitive networks for biomolecules, in which hydrogen bonding, hydrophobic, or ionic contributions will direct recognition. Further developments are expected to have direct impact on applications such as analyte controlled and modulated drug and protein delivery, drug and biological elimination, drug targeting, tissue engineering, and micro- or nano-devices. This work is supported by NSF Grant DGE-99-72770. Copyright © 2003 John Wiley & Sons, Ltd. [source] Biodegradable comb polyesters containing polyelectrolyte backbones facilitate the preparation of nanoparticles with defined surface structure and bioadhesive properties,POLYMERS FOR ADVANCED TECHNOLOGIES, Issue 10-12 2002A. Breitenbach Abstract A major challenge in oral peptide and protein delivery remains the search for suitable carrier systems. Therefore, a new concept was investigated combining a modified three-dimensional architecture, increased hydrophilicity of poly(lactic- co -glycolic acid) (PLGA) and charged groups in a single polymer. Biodegradable comb PLGA were synthesized by grafting short PLGA chains onto different poly(vinyl alcohol) (PVA) based backbone polyols, poly(2-sulfobutyl-vinyl alcohol) and poly(diethylaminoethyl-vinyl alcohol). The polyelectrolyte backbones were obtained by etherification of PVA with charge-containing pendent groups. The comb polymer structure could be confirmed by nuclear magnetic resonance, infrared spectroscopy, differential scanning calorimetry, elemental analysis and measurement of intrinsic viscosity. Nanoparticles (NP), as potential mucosal carriers systems, were prepared by controlled precipitation and investigated as a function of polymer composition. The amphiphilic character and the three-dimensional architecture of the novel polyesters allowed the preparation of small nanoparticles even without the use of surfactants. Surface NMR, surface charge and hydrophobicity determination indicate a core,corona-like NP structure, especially in the case of negatively charged polyesters. A structural model is proposed for the NP with an inner polyester core and an outer charged-groups-containing surface, depending on polymer composition and backbone charge density. The higher the polymer backbone charge density, the more pronounced its influence on the nanoparticle surface properties. The possibility of preparing NP without the use of a surfactant, as well as of designing the NP surface characteristics by polymer backbone charge density and polymer hydrophilic,hydrophobic balance, will be a major advantage in protein adsorption, bioadhesion and organ distribution. This makes these biodegradable polymers promising candidates for colloidal protein and peptide delivery. Copyright © 2003 John Wiley & Sons, Ltd. [source] Protein transduction into human cells by adenovirus dodecahedron using WW domains as universal adaptorsTHE JOURNAL OF GENE MEDICINE, Issue 4 2006A. Garcel Abstract Background Direct protein transduction is a recent technique that involves use of peptide vectors. In this study, we demonstrate that adenovirus dodecahedron (Dd), a virus-like particle devoid of DNA and able to penetrate cells with high efficiency, can be used as a vector for protein delivery. Methods Taking advantage of Dd interaction with structural domains called WW, we have elaborated a universal adaptor to attach a protein of interest to this vector. Results A tandem of three WW structural domains derived from the Nedd4 protein enables the formation of stable complexes with Dd, without impairing its endocytosis efficiency. Our protein of interest fused to the triple WW linker is delivered by the dodecahedron in 100% of cells in culture with on average more than ten million molecules per cell. Conclusion These data demonstrate the great potential of adenovirus dodecahedron in combination with WW domains as a protein transduction vector. Copyright © 2006 John Wiley & Sons, Ltd. [source] Crystallization and preliminary X-ray diffraction studies of the ubiquitin-like (UbL) domain of the human homologue A of Rad23 (hHR23A) proteinACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 9 2009Yu Wai Chen Human homologue A of Rad23 (hHR23A) plays dual roles in DNA repair as well as serving as a shuttle vehicle targeting polyubiquitinated proteins for degradation. Its N-terminal ubiquitin-like (UbL) domain interacts with the 19S proteasomal cap and provides the docking mechanism for protein delivery. Pyramidal crystals of the UbL domain of hHR23A were obtained by the hanging-drop vapour-diffusion method with ammonium sulfate as the crystallizing agent. The crystals diffracted to beyond 2,Ĺ resolution and belonged to the hexagonal space group P6522, with unit-cell parameters a = b = 78.48, c = 63.57,Ĺ. The structure was solved by molecular replacement using the UbL domain of yeast Dsk2 as the search model. [source] A population analysis of VEGF transgene expression and secretionBIOTECHNOLOGY & BIOENGINEERING, Issue 5 2008Golnaz Karoubi Abstract The induction of therapeutic angiogenesis with gene therapy approaches has received considerable interest and some limited clinical success. A major drawback to this approach is a lack of understanding of the pharmacokinetics of therapeutic protein delivery. This has become increasingly more relevant as recent studies have illustrated a defined therapeutic window for angiogenic protein secretion into the local microenvironment. For cell based gene therapies, with cells widely distributed throughout the tissue, this implies that any individual cell must attain a specific secretion rate to produce a local angiogenic response. Here we report a reproducible technique enabling the study of growth factor secretion from individual cells following transient plasmid transfection. We demonstrate significant variability in single cell vascular endothelial growth factor (VEGF) secretion with the majority of total protein secretion arising from a small subpopulation of transfected cells. We demonstrate that VEGF secretion is linearly correlated to intracellular plasmid copy number and protein secretion does not appear to reach saturation within the cell population. The selection of gene therapy approaches that optimize individual cell secretion profiles may be essential for the development of effective gene therapies. Biotechnol. Bioeng. © 2008 Wiley Periodicals, Inc. [source] Three-dimensional fibrous PLGA/HAp composite scaffold for BMP-2 deliveryBIOTECHNOLOGY & BIOENGINEERING, Issue 1 2008Hemin Nie Abstract A protein loaded three-dimensional scaffold can be used for protein delivery and bone tissue regeneration. The main objective of this project was to develop recombinant human bone morphogenetic protein-2 (rhBMP-2) loaded poly(D,L -lactide-co-glycolide)/hydroxylapatite (PLGA/HAp) composite fibrous scaffolds through a promising fabrication technique, electrospinning. In vitro release of BMP-2 from these scaffolds, and the attachment ability and viability of marrow derived messenchymal stem cells (MSCs) in the presence of the scaffolds were investigated. The PLGA/HAp composite scaffolds developed in this study exhibit good morphology and it was observed that HAp nanoparticles were homogeneously dispersed inside PLGA matrix within the scaffold. The composite scaffolds allowed sustained (2,8 weeks) release of BMP-2 whose release rate was accelerated with increasing HAp content. It was also shown that BMP-2 protein successfully maintained its integrity and natural conformations after undergoing the process of electrospinning. Cell culture experiments showed that the encapsulation of HAp could enhance cell attachment to scaffolds and lower cytotoxicity. Biotechnol. Bioeng. 2008;99: 223,234. © 2007 Wiley Periodicals, Inc. [source] | |||||||||||||