High capacity magnetic protein A agarose beads, LOABeads PrtA, were used in the development of a new process for affinity purification of monoclonal antibodies (mAbs) from non-clarified CHO cell broth using a pilot-scale magnetic separator. The LOABeads had a maximum binding capacity of 65 mg/mL and an adsorption capacity of 25-42 mg IgG/mL bead in suspension for an IgG concentration of 1 to 8 g/L. Pilot-scale separation was initially tested in a mAb capture step from 26 L clarified harvest.
View Article and Find Full Text PDFA lab-on-a-chip traveling wave magnetophoresis approach for sensitive and rapid protein detection is reported. In this method, a chip-based magnetic microarray comprising lines of micrometer-sized thin film magnetic elements was used to control the movement of magnetic beads (MBs). The MBs and the chip were functionalized, forming a sandwich-type assay.
View Article and Find Full Text PDFNeural probes are complex devices consisting of metallic (often Pt based) electrodes, spread over an insolating/dielectric backbone. Their functionality is often limited in time because of the formation of scaring tissues around the implantation tracks. Functionalization of the probes surface can be used to limit the glial scar reaction.
View Article and Find Full Text PDFA straightforward, versatile approach to the production of protein gradients on planar and spherical particle surfaces is described. The method is based on the spatially controlled oxidation of thiolated surfaces by Au(III) ions generated via the electrochemical oxidation of a gold electrode in a phosphate-buffered saline solution (10 mM PBS, pH 7.2, 150 mM NaCl).
View Article and Find Full Text PDFHighly dispersed palladium nanoparticles (1-2 nm) supported in large-pore mesocellular foam (MCF; 29 nm) were synthesized. The Pd-nanocatalyst/MCF system was characterized by transmission electron microscopy (TEM), powder X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS). The performance of the Pd nanocatalyst obtained was examined for amine racemization.
View Article and Find Full Text PDFWith the aim of extending the usefulness of an existing biomimetic catalytic system, cobalt porphyrin catalytic units with thiol linkers were heterogenized via chemical grafting to silicon wafers and utilized for the catalytic oxidation of hydroquinone to p-benzoquinone. Atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS) were used to analyze the morphology and composition of the heterogeneous catalyst. The results of the catalytic oxidation of hydroquinone obtained with porphyrins grafted on silicon were compared with those obtained earlier with the same catalyst in homogeneous phase and immobilized on gold.
View Article and Find Full Text PDFExpert Opin Drug Discov
June 2010
Importance Of The Field: Engineering of cell culture substrates provides a unique opportunity for precise control of the cellular microenvironment with both spatial as well as temporal resolutions. This greatly enhances studies of cell-cell, cell-matrix and cell-factor interaction studies in vitro.
Areas Covered In This Review: The technologies used for micropatterning in the biological field over the last decade and new applications in the last few years for dynamic control of surfaces, tissue engineering, drug discovery, cell-cell interactions and stem cell studies are presented.
Brain implants provide exceptional tools to understand and restore cerebral functions. The utility of these devices depends crucially on their biocompatibility and long term viability. We addressed these points by implanting non-functional, NeuroProbes silicon probes, without or with hyaluronic acid (Hya), dextran (Dex), dexamethasone (DexM), Hya+DexM coating, into rat neocortex.
View Article and Find Full Text PDFA novel method of controlled transport of proteins immobilized on micrometre-sized magnetic beads in a lab-on-a-chip environment is presented. Bead motion is controlled by lithographically made magnetic elements forming transportation lines in combination with an applied in-plane rotating magnetic field. In this way, transport of attomole amounts of proteins is controlled with micrometre precision.
View Article and Find Full Text PDFClin Pharmacol Ther
October 2009
The aim of this work was to investigate the pharmacokinetics (PK) of venlafaxine in overdose and the effects of single-dose activated charcoal (SDAC) and whole-bowel irrigation (WBI), alone or in combination, as methods of decontamination. The data included 339 concentration-time points from 76 venlafaxine overdose events (median dose 2,625 (150-13,500 mg)); 69 were slow-release doses. SDAC, WBI, a combination of both, or no decontamination were administered to patients as decided by the treating clinician.
View Article and Find Full Text PDFA novel crossflow filtration methodology is demonstrated for the initial purification of the therapeutic protein, promegapoietin-1a (PMP), produced as inclusion bodies (IBs) in a recombinant Escherichia coli bioprocess. Two strategic separation steps were performed by utilizing a filtration unit with a 1000 kDa polyethersulphone membrane. The first step, aiming for separation of soluble contaminants, resulted in a 50% reduction of the host cell proteins, quantified by total amino acid analysis and a 70% reduction of all DNA, quantified by fluorometry, when washing the particulate material with a 10mM EDTA in 50mM phosphate buffer, pH 8.
View Article and Find Full Text PDFA great challenge in functional or interaction proteomics is to map protein networks and establish a functional relationship between expressed proteins and their effects on cellular processes. These cellular processes can be studied by characterizing binding partners to a "bait" protein against a complex background of other molecules present in cells, tissues, or biological fluids. This so-called ligand fishing process can be performed by combining surface plasmon resonance biosensors with MS.
View Article and Find Full Text PDFJ Colloid Interface Sci
September 2005
In this investigation, the structure, stability, and orientation of bovine serum albumin (BSA) adsorbed onto silica particles were studied using differential scanning calorimetry (DSC) and limited proteolysis in combination with mass spectrometry (MS). DSC gave information on the overall structural stability of BSA while limited proteolysis was used to probe the accessibility of enzymatic cleavage sites, thereby yielding information on the orientation and structure of BSA adsorbed to silica surfaces. Thermal investigation of BSA in various buffers, both free in solution and in the adsorbed state, showed that solutes that surround the protein played an important role with respect to the overall structural stability and the structural heterogeneity of BSA.
View Article and Find Full Text PDFMicrocontact printing is a remarkable surface patterning technique. Developed about 10 years ago, it has triggered enormous interest from the surface science community, as well as from engineers and biologists. The last five years have been rich in improvements to the microcontact printing process itself, as well as in new technical innovations, many designed to suit new applications.
View Article and Find Full Text PDFJ Mater Sci Mater Med
March 2004
Fluorinated hydroxyapatite is known to be less soluble by body fluids, resulting in enhanced resistance to biodegradation in vivo conditions, as compared to the pure hydroxyapatite ceramics. The present work was aimed at the investigation of the effect of minor additions of ultrafine fluorapatite (up to 10 wt%) on the sintering behavior and mechanical properties of hydroxyapatite ceramics. In vitro testing for the osteoblast-like cells viability and proliferation was performed with the samples of varying fluorapatite content.
View Article and Find Full Text PDFThe structural stability of hen egg white lysozyme in solution and adsorbed to small colloidal silica particles at various surface concentrations was investigated using hydrogen-deuterium (H/D) exchange in combination with mass spectrometry (HDX-MS) and differential scanning calorimetry (DSC). The combination of HDX-MS and DSC allows a full thermodynamic analysis of the lysozyme structure as both the enthalpy and the Gibbs free energy can be derived from the various measurements. Moreover, both HDX-MS and DSC provide information on the relative structural heterogeneity of lysozyme in the adsorbed state compared to that in solution.
View Article and Find Full Text PDFNon-viral gene therapy constitutes an alternative to the more common use of viral-mediated gene transfer. Most gene transfer methods using naked DNA are based upon non-sequence-specific interactions between the nucleic acid and cationic lipids (lipoplex) or polymers (polyplex). We have developed a technology in which functional entities hybridize in a sequence-specific manner to the nucleic acid (bioplex).
View Article and Find Full Text PDFA new method is presented for monitoring the conformational stability of various parts of a protein that is physically adsorbed onto nanometer-sized silica particles. The method employs hydrogen/deuterium (H/D) exchange of amide hydrogens, a process that is extremely sensitive to structural features of proteins. The resulting mass increase is analyzed with Fourier transform ion cyclotron resonance (FTICR) mass spectrometry.
View Article and Find Full Text PDFJ Colloid Interface Sci
October 2002
A novel method to derivatize silicon surfaces with 3-mercaptopropylsilane molecules has been developed and optimized. This method is based on an argon flow that increases the evaporation rate of the silane molecules by lowering the partial pressure of the silane molecules in gas phase above the liquid silane, at room temperature. X-ray photoelectron spectroscopy studies of the surfaces showed a dense monolayer coverage as well as hydrolysis of the silane methoxy groups.
View Article and Find Full Text PDFHuman plasma fibronectin (Fn) is a large flexible protein stabilized by intermolecular ionic interactions forming a compact structure. On altering solution conditions, the structure can revert to a more expanded state, thereby exposing previously hidden domains (e.g.
View Article and Find Full Text PDFRecent research suggests that the biocompatibility of an implant is to a large extent determined by selective adsorption of proteins from surrounding body fluids. Protein adsorption from human plasma onto two bioactive glass-ceramics (RKKP and AP40) which differ in La and Ta content, was studied by means of chromatography and two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). The quantitative analysis showed that the glass-ceramics have good protein binding capacities indicating multilayer formation.
View Article and Find Full Text PDFThe protein/biomaterial interactions of three biomaterials used in hard tissue surgery were studied in vitro. A dynamic flow system and two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) were used to investigate the adsorption of proteins from diluted human plasma on hydroxyapatite, alumina and zirconia, with regard to total protein binding capacity, relative binding capacity for specific proteins and flow-through and desorption patterns. The ceramics were characterized regarding physicochemical properties; namely, chemical composition by elementary analyses and specific surface, pore volume and pore size distribution using the BET-method and Hg-porosimetry.
View Article and Find Full Text PDFJ Colloid Interface Sci
May 2001
Electrostatic effects on protein adsorption were investigated using differential scanning calorimetry (DSC) and adsorption isotherms. The thermal denaturation of lysozyme, ribonuclease A (RNase), and alpha-lactalbumin in solution and adsorbed onto silica nanoparticles was examined at three concentrations of cations: 10 and 100 mM of sodium and 100 mM of sodium to which 10 mM of calcium was added. The parameters investigated were the denaturation enthalpy (DeltaH), the temperature at which the denaturation transition was half-completed (T(m)), and the temperature range of the denaturation transition.
View Article and Find Full Text PDFJ Am Soc Mass Spectrom
April 2001
The isotopic exchange of amide hydrogens in proteins in solution strongly depends on the surrounding protein structure, thereby allowing structural studies of proteins by mass spectrometry. However, during electrospray ionization (ESI), gas phase processes may scramble or deplete the isotopic information. These processes have been investigated by on-line monitoring of the exchange of labile deuterium atoms in homopeptides with hydrogens from a solvent suitable for ESI.
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