Abstract: A test device having a micro flow channel including a reaction part where a reactant that is reactive to a tested chemical dispersed in a tested fluid is fixed, and at least one actuator for actuating the tested fluid to move in at least one of two opposite sides of the micro flow channel so as to homogenize a density distribution of the tested chemical in the tested fluid. The tested fluid is sent in the micro flow channel a plurality of times.

Abstract: Isoelectric focusing devices configured for multiplex separation of sample components of interest in a polymeric separation medium are provided. Also provided are methods of using the devices as well as systems and kits that include the devices. The devices, systems and methods find use in a variety of different applications, including diagnostic and validation assays.

Abstract: The invention provides a dry electroblotting system for dry blotting gels, in which the system includes an electroblotting transfer stack that comprises an analysis gel and a blotting membrane, an anode, a body of anodic gel matrix juxtaposed with the anode between the anode and the transfer stack, a cathode, and a body of cathodic gel matrix juxtaposed with the cathode between the cathode and the transfer stack, in which the anodic gel matrix and the cathodic gel matrix each comprise an ion source for electrophoretic transfer. The dry electroblotting system does not use any liquid buffers that are added to the system just before electroblotting (such as when the transfer stack is being assembled). The anode, the cathode, or both can be separate from a power supply and provided as part of a disposable electrode assembly that also includes a body of gel matrix that includes ions for electrophoretic transfer.

Abstract: The present invention relates to a method for purifying darbepoetin alfa by selectively separating only a structural isoform having a high content of sialic acid from a mixture of structural isoforms of darbepoetin alfa having various contents of sialic acid. Since the method of the present invention is a novel method for purifying darbepoetin alfa which can be conveniently and simply produced, it is possible to remarkably increase productivity due to process efficiency improvement, as well as to yield high purity darbepoetin alfa when mass-producing darbepoetin alfa according to the present invention.

Abstract: The present invention discloses a gel electrophoresis chip, comprising a first substrate, a first plurality of parallel gel strips formed on the first substrate, respectively extending along a first direction and having a certain width; and a second plurality of isolation segments formed on the first substrate, respectively located between adjacent gel strips and extending along a second direction different from the first direction, the isolation segments being arranged to form a microwell array together with the gel strips. After the gel electrophoresis chip achieves conventional protein two-dimensional gel electrophoretic separation, protein samples suitable for mass spectrometry analysis are prepared in high throughput, thus greatly reducing the pretreatment time of mass spectrometry analysis, thereby being suitable for proteomic analysis of biological samples.

Abstract: Microfluidic devices and methods for using the same are provided. Embodiments include microfluidic devices that have a first separation region configured to separate a sample along a first directional axis based on a first property, and a second separation region in fluid communication with the first separation region and configured to separate the sample along a second directional axis based on a second property. Also provided are methods of using the devices as well as systems and kits that include the devices. The devices, systems and methods find use in a variety of different applications, including diagnostic and validation assays.

Abstract: A clearing agent and mounting solution for microscopy is disclosed comprising (a) trichloroethanol, (b) optionally, trichloroacetic acid, (c) optionally, glycerol and (d) optionally, water, where the refractive index of the solution is greater than or equal to about 1.3810. The solution can further comprise a C1-C6 alcohol, other acids, and/or a stain. The solution can also comprise derivatives and/or analogs of 2,2,2-trichloroethanol and/or trichloroacetic acid. Also disclosed is a method of preparing specimens for microscopy comprising (a) applying a specimen to a microscope slide or a cuvette, (b) applying a quantity of the above solution sufficient to mount the specimen, and (c) optionally applying a cover slip. The solution can be used effectively with stains or dyes, and with fresh, partially dry or dried materials, and for temporary or semi-permanent to permanent mounting.

Abstract: A system and method for generating a digital image in fluorescence gel imaging is disclosed. The method includes providing a gel sample and placing the gel sample on a flat panel detector having array of photodiodes and transistors that collect light generated from the gel sample. The gel sample is illuminated using a light source integrated into the flat panel imaging system and light emitted by the gel sample responsive to an excitation of the gel sample by light provided by the light source is then collected, with the light emitted by the gel sample being collected by the array of photodiodes of the flat panel detector and converted to electric charges to generate light data. The light data is then processed to generate a digital image of the gel sample.

Abstract: Multi-directional microfluidic devices and methods for using the same are provided. Aspects of the invention include microfluidic devices that are configured to subject a sample to two or more directionally distinct flow fields, and include a separation medium and a binding medium, where the binding medium is in fluid communication with the separation medium. Also provided are methods of using the devices as well as systems and kits that include the devices. The devices, systems and methods find use in a variety of different applications, including diagnostic and validation assays.

Abstract: A device for isoelectric focusing. The device comprises a focusing container configured to contain an electrolyte solution and having a longitudinal axis and at least one electrolysis unit mounted in a close proximity to the longitudinal axis. Each electrolysis unit injects an ion flow into the focusing container so as to create a pH gradient having a plurality of steps in the electrolyte solution, along the longitudinal axis. Each step has a substantially uniform pH level and the pH gradient is defined by at least one pH ramp between every two sequential steps of the plurality of steps.

Abstract: The present invention provides a mechanism for separating or isolating charged particles under the influence of an electric field without metal electrodes being in direct contact with the sample solution. The metal electrodes normally in contact with the sample are replaced with high conductivity fluid electrodes situated parallel and adjacent to the sample. When the fluid electrodes transmit the electric field across the sample, particles within the sample migrate according to their electrophoretic mobility.

Abstract: Provided is a device comprising an upper chamber, a middle chamber and a lower chamber, wherein the upper chamber is in communication with the middle chamber through a first pore, and the middle chamber is in communication with the lower chamber through a second pore, wherein the first pore and second pore are about 1 nm to about 100 nm in diameter, and are about 10 nm to about 1000 nm apart from each other, and wherein each of the chambers comprises an electrode for connecting to a power supply. Methods of using the device are also provided, in particular for sequencing a polynucleotide.

Abstract: Methods and devices for purifying, detecting, and collecting analytes fractionated based on pI, separating analytes via electrophoresis and pI, and purifying a target molecule using pI focusing and subsequent crystallization are provided.

Abstract: A method of separating a mixture of a plurality of molecular analytes having different isoelectric points (pIs). The method comprises placing a solution containing a mixture of a plurality of molecular analytes in a separation volume, generating a pH profile having a plurality of pH zones across an axis of the separation volume, and adjusting a profile of the pH profile to induce a migration of a first molecular analyte along the axis apart from a second molecular analyte. The first and second molecular analytes having different pIs.

Abstract: The present invention relates to the field of electrophoresis and more specifically to a gel or strip for separating peptide components by isoelectric focusing by producing IPG (immobilized pH gradient) gels or strips in a novel way before focusing. More closely, the peptides are focused in a novel IPG gel including an uneven or non-linear pH gradient having at least three separate stepwise arranged pH-intervals. After focusing the peptide resolution is high and the peptides are evenly distributed along the gel.

Abstract: An improved method of separating a macromolecule by isoelectric focusing comprising subjecting the macromolecule to electrophoresis in an isoelectric-focusing medium including a substantially thiol-free reducing agent, preferably a trivalent phosphorous compound and more preferably tributyl phosphine, the improvement being the solubility and focusing of the macromolecule is enhanced compared to isoelectric focusing of the same macromolecule in a similar isoelectric-focusing medium containing a thiol-reducing agent.

Abstract: A method and apparatus for continuously separating or concentrating molecules that includes flowing two fluids in laminar flow through an electrical field and capturing at one of three outputs a fluid stream having a different concentration of molecules.

Abstract: Provided are a method and apparatus for measuring an isoelectric point using a field effect transistor. The method includes providing a field effect transistor including a substrate, source and drain electrodes disposed on the substrate and spaced apart from each other, and a channel region between the source and drain electrodes, providing a first electrolyte solution having a first concentration to the channel region of the field effect transistor and measuring a first current value of the channel region between the source and drain electrodes, providing a second electrolyte solution having a second concentration greater than the first concentration and measuring a second current value of the channel region between the source and drain electrodes, and determining the isoelectric point of the field effect transistor or a material disposed on the field effect transistor using a difference between the first and second current values.

Abstract: The present invention provides methods, compositions, and kits for protein crystallization. The present invention involves electrophoretically focusing at least a first protein species within a matrix comprising at least 2 regions of different pH, the protein being present in amount sufficient to permit crystallization within said pH gradient.

Abstract: The present disclosure relates to novel humanized antibodies able to inhibit tumor growth, as well as the amino and nucleic acid sequences coding for such antibodies. From one aspect, the disclosure relates to novel anti-JAM-A homogeneous humanized antibodies able to inhibit tumor growth. The disclosure also comprises the use of such antibodies as a drug for the preventive and/or therapeutic treatment of cancers, as well as compositions comprising such antibodies in combination with other anticancer compounds. The disclosure also relates to a method for the preparation of such novel humanized antibodies.

Abstract: A nanoparticle translocation device includes a first reservoir having a first reservoir electrode, a second reservoir having a second reservoir electrode, and at least one nanopore providing fluid communication between the first and second reservoirs. The device also includes one or more inner electrode portions on an inner wall of the nanopore and one or more outer electrode portions disposed on an outer wall of the nanopore. The device further includes at least one DC voltage supply for selectively applying a DC voltage to each of the first reservoir electrode, the second reservoir electrode, and the outer electrode layer, where the inner electrode portions, the outer electrode portions, and the nanopore are in a substantially coaxial arrangement.

Abstract: An electrophoresis gel assembly comprising a gel slab, at least two sample wells and at least one focusing buffer well arranged between two adjacent sample wells and wherein focusing barriers are arranged between the focusing buffer well and the sample wells. A high conductivity focusing buffer can be applied to the focusing buffer well to obtain a lateral focusing of the sample bands during electrophoresis.

Abstract: Methods and devices for detecting and collection analytes fractionated based on pI, separating analytes via electrophoresis and pI, and purifying a target molecule using pI focusing and subsequent crystallization are provided.

Abstract: The present invention provides methods, compositions, and kits for protein crystallization. The present invention involves electrophoretically focusing at least a first protein species within a matrix comprising at least 2 regions of different pH, the protein being present in amount sufficient to permit crystallization within said pH gradient.

Abstract: The invention provides methods for detecting an interaction between an analyte and a biomolecule. The method comprises separating at least one biomolecule according to its isoelectric point in the presence of a given analyte and detecting an interaction between the analyte and a biomolecule using fluorescence anisotropy. The method may further comprise collecting the analyte-biomolecule complex and analyzing the biomolecule.

Abstract: The present invention generally relates to devices and methods for the separation of species, including biological species. Some embodiments involve the use of free flow isoelectric focusing (FF-IEF) devices in the separation of a mixture of species. Various device configurations and/or features may enhance the performance of the devices, providing faster and more efficient devices and methods for separating mixtures of species with high resolution. In some embodiments, separation of a mixture of species may be achieved with high resolution and with short sample residence times. Such devices may provide simplified, inexpensive, and optionally disposable devices for the separation of chemical and biological species and may optionally be integrated with orthogonal separation techniques, such as SDS-PAGE or capillary electrophoresis.

Abstract: The present invention contemplates various devices that are configured to separate a sample, which contains more than one unique species, into any desired number of sub-samples by passing the sample across a like number of separation media configured for a first separation protocol. Each of the sub-samples may be further separated by an additional separation protocol, thereby creating a plurality of mini-samples, each of which may be further separated and/or analyzed. The invention also contemplates using a simple method of using conduits to form a fluid path that passes through a plurality of separation media, each of which media is configured to isolate a particular sub-sample. After various sub-samples of the sample are isolated by the various separation media, the conduits may be removed, thereby enabling each of the isolated sub-samples to be further separated and/or analyzed independent of any other sub-sample.

Abstract: Provided is a multi-channel apparatus for non-gel based two-dimensional protein separation. One or more flat channels are arranged in parallel and have an isoelectric focusing section for primarily separating proteins from protein samples according to isoelectric point (pI) and a flow field-flow fractionation section for secondarily separating the primarily separated proteins according to molecular weight, thereby making it possible to simultaneously separate the proteins in multiple channels, to increase a protein separation speed, and to overcome limitation to sample injection due to an increase in channel volume. The apparatus can separate the proteins according to pI and molecular weight, be safe from denaturation of the protein in the process of protein separation, automatically remove an ampholyte used for pI-based separation, and separately detecting the separated proteins to identify the proteins using nanoflow liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS-MS).

Abstract: The procedures involved in isoelectric focusing that entail the use of an electric current can be performed with the gel side of the IPG strip facing either up or down in a tray and electrode assembly in which the electrodes are mounted on an insert from which the inner ends of the electrodes extend into the troughs to reside either below the IPG strips or above them. Slots in one part (either the insert or the tray) mate with walls or partitions in the other part to secure the two parts together, and the heights of the electrodes within the troughs of the tray can be adjusted by the height of the insert in the tray or by a resilient mounting of the electrode to the insert.

Abstract: An ensemble of components and methods are disclosed for utilizing two-dimensional electrophoresis in polyacrylamide or related polymer gels using common, existing and familiar electrophoresis formats and equipment. The disclosed invention makes two-dimensional electrophoresis convenient and easy to use for individuals already using vertical “mini-gel” type systems. The invention discloses the combination of pre-cast disposable gels for both first and second separation dimensions using novel support devices and cassettes that simplify the difficult multiple sample handling and processing steps Inherent in ordinary two-dimensional electrophoresis methods. Devices and methods are disclosed in said invention that provide exceptional gel to gel reproducibility.

Abstract: The present invention relates to the field of electrophoresis and more specifically to a gel or strip for separating peptide components by isoelectric focusing by producing IPG (immobilised pH gradient) gels or strips in a novel way before focusing. More closely, the peptides are focused in a novel IPG gel including an uneven or non-linear pH gradient having at least three separate stepwise arranged pH-intervals. After focusing the peptide resolution is high and the peptides are evenly distributed along the gel.

Abstract: Described herein is a sample preparation device including a sample delivery source, an inline means of transferring the sample from the sample source into a deformable channel within a pressure vessel, and out of the channel into downstream analysis components, a deformable channel disposed within the pressure vessel, the deformable channel having an inlet end and an outlet end fluidly connectable to high pressure valves and a means to measure the fluid pressure within the deformable channel, an external source of a controlled pressurized fluid fluidly connectable to the pressure vessel and a controller system that monitors and controls the sample fluid pressure by control of the external pressure vessel fluid.

Abstract: An improved method of separating a macromolecule by isoelectric focusing comprising subjecting the macromolecule to electrophoresis in an isoelectric-focusing medium including a substantially thiol-free reducing agent, preferably a trivalent phosphorous compound and more preferably tributyl phosphine, the improvement being the solubility and focusing of the macromolecule is enhanced compared to isoelectric focusing of the same macromolecule in a similar isoelectric-focusing medium containing a thiol-reducing agent.

Abstract: The invention includes nanochannel devices and methods for using such nanochannel devices for separating molecules, ions and biomolecules. The nanochannel devices have at least one nanochannel through which fluid can move, wherein ionic double layers form in the fluid near walls of the nanochannel and those ionic double layers overlap within the nanochannel. Electrical voltage can be applied to the nanochannel to modify an electrostatic potential in the nanochannel and thereby control movement of ions and biomolecules through the nanochannel. The invention also includes arrays and networks of such nanochannel devices.

Abstract: Devices are provided for determining the isoelectric point of a charged analyte, comprising a titration chamber and an electrode chamber. The electrode chamber comprises at least two electrodes, for example, an electrode array. Either or both of the titration chamber and the electrode chamber may have a shaped geometry. The electrodes are operative, in conjunction with the shaped geometry of the chamber(s) where appropriate, to generate an electric field gradient in the titration chamber. Permeable material separates the titration chamber and the electrode chamber. A pH Sensor is located in the titration chamber for obtaining the pH of the first fluid. Certain preferred embodiments further include an analyte band detector for detecting the presence and optionally the location of a focused band of charged solute.

Abstract: The invention provides methods and compositions for determining whether a subject is at risk of developing age-related macular degeneration, for example, the wet or neovascular form of age-related macular degeneration. The method involves determining whether the subject has a protective variant and/or a risk variant at a polymorphic site in the HTRA1 gene. In addition, the invention provides a method of treating or slowing the progression of age-related macular degeneration by reducing the expression of the HTRA1 gene, or reducing the biological activity of the HTRA1 gene product.

Abstract: The invention relates to a method of screening a subject for at least one risk factor associated with a neurodegenerative disease such as Alzheimer's disease comprising detecting the presence or absence of at least one risk marker in the HtrA1 gene (PRSS11). Furthermore, diagnostic kits as well as therapeutic approaches are provided.

Abstract: The present invention is related to a method of separation of compounds by electrophoresis in which the compounds such as genes, proteins, etc. may be analyzed very precisely as samples are introduced directly into the separation tubes of the chip at the collection site, and therefore, it is not necessary to have separate fluid paths or individual sample storing apparatus that have been necessary for the conventional electrophoresis; it is easy to make the chips as the structure of the chip becomes extremely simple, and high-density arrangement of the separation tubes is enabled; and further, the compounds such as genes, proteins, etc. may be analyzed very precisely without interference in the storage tubs by using a non-polar solvent as the solvent of the sample storage tub.

Abstract: The present invention relates to an electrode bridge or sample application bridge for use in electrophoresis, preferably isoelectric focusing, IEF. More closely, the invention relates to an electrodic bridge or sample loading bridge for preventing denaturant depletion from the IPG (immobilized pH gradient) gel and, optionally, for loading samples onto the IPG gel. The pH of the bridge is adjustable for adoption to positioning at the acid as well as basic end of the IPG strips.

Abstract: The present invention is to provide a sample separation instrument used in a sample separation apparatus which includes: holding means for holding a first medium supporter which supports a first medium; and driving means for moving fixing means or the holding means in a direction parallel or perpendicular to a plane whose sides extend in the first direction and in the second directions. The sample separation instrument includes an insulator for storing a second medium which allows a sample separated in the first medium in the first direction to be further separated in the second direction different from the first direction, wherein: the insulator includes a first opening and a second opening each of which defines the second direction in which the second medium is electrified, and the second opening has a shape which allows the first medium containing the separated sample to be attached to the second medium along the first direction.

Abstract: The present invention generally relates to compositions comprising the proteose peptone fraction (PPf). In particular, the present invention relates to a method for the production of an extract comprising a demineralised protein fraction depleted in ?-lactoglobulin and enriched in the PPf and to uses of these extracts, e.g. in a food product, a food supplement, a nutritional, a pharmaceutical and/or a cosmetic composition, for example as emulsifier or as foaming agent. The PPf fraction of the present invention may be obtained by adjustment of the pH of an aqueous native protein dispersion to about 5.6 to 8.4, or to about 3.5 to 5.0, heating the aqueous native protein dispersion to about 70-95° C. for about 10 seconds to 60 minutes, removing at least a part of the formed solid large molecular weight aggregates with a diameter of at least 100 nm from the aqueous protein dispersion after heating and collecting the remaining liquid fraction of the dispersion.

Abstract: The invention provides methods for detecting an interaction between an analyte and a biomolecule. The method comprises separating at least one biomolecule according to its isoelectric point in the presence of a given analyte and detecting an interaction between the analyte and a biomolecule using fluorescence anisotropy. The method may further comprise collecting the analyte-biomolecule complex and analyzing the biomolecule.

Abstract: A device providing an arrangement for the separation of a sample mixture for analytical reasons based on multidimensional gel electrophoresis and method thereof are disclosed. The separation involves a first separation in a first dimension of the device on the basis of isoelectric points and a second separation in a second dimension of the device on the basis of molecular size. At least two gel strips for the first dimension separation step and a corresponding gel for the second dimension separation step are provided. The two gel strips are arranged on a single carrier either on the same side or on opposite sides. By having at least two gel strips arranged in such a manner at least two analytical processes can be executed in parallel on the single carrier without the use of valves.

Abstract: A method for protein identification in complex mixtures that utilizes affinity selection of constituent proteolytic peptide fragments unique to a protein analyte. These “signature peptides” function as analytical surrogates. Mass spectrometric analysis of the proteolyzed mixture permits identification of a protein in a complex sample without purifying the protein or obtaining its composite peptide signature.

Abstract: The present invention provides a novel and advantageous method for separating analytes by free flow electrophoresis. The methods are particularly suitable for depleting major constituents such as albumin from samples of biological origin, optionally combined with a further separation of the remaining portion of the sample. The sample portions recovered from the method can be used advantageously in downstream applications such as 1D or 2D-PAGE, HPLC or mass spectrometric analysis. Also provided are buffer systems, kits comprising such buffer systems, and devices for carrying out the free flow electrophoretic separation methods of the present invention.

Abstract: The procedures involved in isoelectric focusing that entail the use of an electric current can be performed with the gel side of the IPG strip facing either up or down in a tray and electrode assembly in which the electrodes are mounted on an insert from which the inner ends of the electrodes extend into the troughs to reside either below the IPG strips or above them. Slots in one part (either the insert or the tray) mate with walls or partitions in the other part to secure the two parts together, and the heights of the electrodes within the troughs of the tray can be adjusted by the height of the insert in the tray or by a resilient mounting of the electrode to the insert.

Abstract: The invention relates to a method for the diagnostic investigation of biological samples from a person for inflammation of the liver, in particular hepatic fibrosis and/or cirrhosis of the liver, where the sample is investigated for one or more proteins as markers of inflammation of the liver, in particular hepatic fibrosis and/or cirrhosis of the liver, where a concentration of the proteins which is elevated or decreased by comparison with the healthy state indicates the presence of an inflammation of the liver, in particular a hepatic fibrosis and/or cirrhosis of the liver.

Abstract: The present invention relates to methods and devices for forming a plurality of wells on a gel containing an analyte. The invention further provides a system for eluting a liquid analyte reagent mixture from a gel. The invention is useful in the separation of biological molecules such as nucleic acids, carbohydrates, proteins and peptides. In particular, the invention has utility for separating and eluting peptides from isoelectric focusing gels.

Abstract: The present invention relates to the field of isoelectric focussing or 2D electrophoresis, in particular isoelectric point markers with fluorescence detection. The marker comprises an oligopeptide covalently labelled with a fluorescent dye and is characterised in that the dye has a net charge that will maintain the overall net charge of the oligopeptide upon being bound to the oligopeptide. The invention also relates to an electrophoretic method for analysis of proteins using these markers.

Abstract: Provided herein are rapidly-rehydratable prior-cast, dehydrated, electrophoresis separation media particularly useful for isoelectric focusing, including methods of making, and methods of use thereof.