1,721,099 research outputs found
Sol-gel derived silica polymers for the sustained release of proteins
No full textThe use of wet silica based polymers obtained by the sol-gel technology for the entrapment and sustained release of macromolecular bioactive compounds and, more specifically, proteins with pharmaceutical applications, is disclosed. The wet-gel formulations are obtained by a method which comprises mixing an alkoxysilane solution, a gelling catalyst and a buffered water solution of a biologically active principle and subsequently casting the thus-obtained mixture into suitable containers without previous or subsequent partial or complete removal of the water solvent; the containers are then sealed after casting in order to preserve the wet state of the gel. The wet-gel formulations may then be used for the manufacture of a ready-to-use injectable solution by mixing the silica wet-gel formulation with water or with a physiological water solution
Synthesis and optimization of sensing layer for Grating-Coupled Surface Plasmon Resonance detection of proteins and nucleic acids
No full textBioconjugation in pharmaceutical chemistry
No full textPolymer conjugation is of increasing interest in pharmaceutical chemistry for delivering drugs of simple structure or complex compounds such peptides, enzymes and oligonucleotides. For long time drugs, mainly with antitumoral activity, have been coupled to natural or synthetic polymers with the purpose of increasing their blood permanence time, taking advantage of the increased mass that reduces kidney ultrafiltration. However only recently complex constructs were devised that exploit the ‘enhanced permeability and retention’ (EPR) effect for an efficient tumor targeting, the high molecular weight for adsorption or receptor mediated endocytosis and finally a lysosomotropic targeting, taking advantage of acid labile bonds or cathepsin susceptible polypeptide spacers between polymer and drug. New original, very active conjugates of this type, as those based on poly(hydroxyacrylate) polymers, are already in advanced state of development. Labile oligonucleotides, including antisense drugs, were also successfully coupled to polymers in view of an increased cell penetration and stabilization towards nucleases. However, the most active research activity resides in the field of polypeptides and proteins delivery, mainly for the two following reasons: first of all because a great number of therapeutically interesting compounds are now being produced by genetic engineering in large quantity and, secondly, because these products are difficult to administer to patients for several inherent drawbacks. Proteins are in fact easily digested by many endo- and exo-peptidases present in blood or in other body districts; most of them are immunogenic to some extent and, finally, they are rapidly excreted by kidney ultrafiltration. Covalent polymer conjugation at protein surface was demonstrated to reduce or eliminate these problems, since the bound polymer behaves like a shield hindering the approach of proteolytic enzymes, antibodies, or antigen processing cell. Furthermore, the increase of the molecular weight of the conjugate allows to overcome the kidney elimination threshold. Many successful results were already obtained in peptides and proteins, conjugated mainly to water soluble or amphiphilic polymers like poly(ethylene glycol) (PEG), dextrans, or styrene–maleic acid anhydride. Among the most successful are the conjugates of asparaginase, interleukin-2 or -6 and neocarcinostatin, to remind some antitumor agents, adenosine deaminase employed in a genetic desease treatment, superoxide dismutase as scavenger of toxic radicals, hemoglobin as oxygen carrier and urokinase and streptokinase as proteins with antithrombotic activity. In pharmaceutical chemistry the conjugation with polymers is also of great importance for synthetic applications since many enzymes without loss of catalytic activity become soluble in organic solvents where many drug precursors are. The various and often difficult chemical problems encountered in conjugation of so many different products prompted the development of many synthetic procedures, all characterized by high specificity and mild condition of reaction, now known as ‘bioconjugation chemistry’. Bioconjugation developed also the design of new tailor-made polymers with the wanted molecular weight, shape, structure and with the functional groups needed for coupling at the wanted positions in the chain
Conjugates of Peptides and Proteins to Polyethylene Glycols
No full textThis chapter provides a critical overview of the technology presently available in the field of protein PEGylation. The chemistry of the polymer and of its reactive derivatives is discussed and presented together with several protocols used to obtain PEG-protein conjugates. The coupling protocols are critically discussed on the basis of the properties of the protein to be modified and those desired for the final product. Methods for product purification and characterization are also provided. The overall information provided will guide the reader toward all of the critical steps involved in the preparation of PEG-protein adducts
MICROFLUIDIC DEVICES AND METHODS OF GENERATING AND USING SAME
No full textA microfluidic device comprising a substrate having formed therein microfluidic paths, at least a portion of the microfluidic paths having attached thereto a plurality of monolayers, wherein at least a portion of the monolayers comprises a photoactivatable group capable of generating a reactive group upon exposure to a light source, the reactive group being for binding a screenable moiety
An approach to increased polyplex gene delivery by peptides selected from a phage display library
No full textPhage display libraries were screened for peptides to be incorporated in nonviral gene delivery vehicles. Cells in culture were incubated with heptamer random peptide libraries displayed on M13 bacteriophages in three to five copies per phage. Surface-adherent phages were removed or inactivated and the cells were fractionated in a nuclear pellet and supernatant. Bacteriophages from each of the two fractions were amplified and reincubated with the cells. Three successive rounds of selection were performed. Eighteen sequenced clones revealed 14 different sequences. Two sequences were homologous to segments of the HIV gp120 protein. For three sequences, the corresponding synthetic peptides were generated and attached via avidin-biotin to polylysine-condensed plasmid DNA containing a reporter gene. The addition of the peptides led to 8-14 times increase in the expression of the reporter
Optimized avidin nucleic acid nanoassemblies by a tailored PEGylation strategy and their application as molecular amplifiers in detection
Full text linkAvidin was recently found to display the ability to interact with high affinity with nucleic acids. In this work, we investigated how this property is affected by the protein modification with poly(ethylene glycol) (PEG). More precisely, we studied the influence of the size and geometry of the polymer and of the mode of anchorage to the protein surface. To this end, we synthesized five PEG derivatives capable of PEGylating avidin either through covalent attachment to its lysine primary amines or by exploiting its biotin binding pockets. Several differently PEGylated avidin derivatives were then obtained, which were later tested for their affinity for plasmid DNA by means of the electrophoretic mobility assay. The results show that covalent PEGylation reduces the affinity for DNA in a dose-dependent manner, whereas PEG anchoring through the biotin binding sites does not, even when bulky and high MW biotin-PEG derivatives are used. We then investigated how the size and molecular weight of the biotin-PEG affects the solubility and stability of avidin-nucleic acid nanoassemblies in physiological buffer. Among the biotin-PEG derivatives synthesized in this work, the branched forms were more efficient in protecting particle surface and preventing their aggregation. Full nanoparticle solubility was achieved by saturating 30% of the biotin binding sites with a 2 x 5 kDa branched derivative. the optimized avidin nucleic acid nanoassemblies (ANANAS) were employed in a model analytical test where they showed at least 40-fold higher efficiency than monomeric avidin in recognizing biotinylated surface immobilized IgGs. The results pave the way toward the application of this novel nanosystem in biomedicine
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