University Patents, Inc.

Protein Technologies, Inc.

Other

The United States of America As Represented by the Secretary of

Milan Bier

Terry D Long

Anthony R Ford

Leon E Barstow

Rodolfo Bacheleir Marquez

Garland E Twitty

James A Ostrem

Peter Reynolds Foster

Rodolfo B Marguez

German Eduardo Fuentes

An elastomeric cell is used as the disposable part of an apparatus for isoelectric focusing in free solution (without gels) in the 0.5 to 5 ml volume range. An inlet port is used for priming the cell and end-connectors for coupling with electrodes. A grid of parallel rods compresses the cell against a cold plate, thereby causing swelling of the skin of the cell between pairs of rods and forming contiguous fluid bubble-compartments for IEF separation. Before collection of separated fractions, the gap between the rods and the plate is further reduced so as to create distinct fluid compartments which now contain discrete products of separation. The separated fractions are collected by syringe-like devices by puncturing the elastomer skin. The plate, rods and deformable cell are capable of rotation or gentle rocking motion around the cell's main axis to avoid gravitational convection during the focusing process.

Variable-volume disposable isoelectric focusing cell and method of isoelectric focusing

The object of the present invention is the oral administration of animal immunoglobulins for the treatment of gastrointestinal disorders caused by bacterial and/or yeast overgrowth. Immunoglobulins derived from the blood, plasma or serum of animals, such as cow, goats, sheep and pigs, contain a broad spectrum of antibodies to bacteria and yeast that afflict the gastrointestinal tract of human patients.

Oral immunotherapy of bacterial overgrowth

The objective of combinatorial chemical synthesis is the preparation of a very large number of different molecular entities by a randomized process of sequential chemical syntheses. The present invention provides a combinatorial vessel comprising a reaction region and a mixing region, adjacent to each other, which can be used to practice combinatorial chemical synthesis. The reaction region comprises a plurality of reaction cavities, in which a plurality of different chemical reactions may be performed simultaneously on resin beads. The resin beads are then transferred to the mixing region and mixed into a single group. Then, the resin beads are transferred into the reaction cavities. The process of performing reactions, transferring to the mixing region, mixing, and transferring to the reaction region can be repeated multiple times to obtain a large number of resin beads, each of which has been exposed to a series of reactions different from most of the other resin beads.

Apparatus and process for conducting combinational chemical syntheses

A multi-channel separation device is provided for use in separation technology, particularly in electrophoresis and in isoelectric focusing. The device includes two mating plates, at least one of which contains an array of at least three cavities arranged to define a serpentine fluid pathway through the cavities when the plates are mated. Each of the cavities includes fluid input and output ports to facilitate transfer of process fluid through the cavities. A septum is disposed between the plates for streamlining fluid flow between the input and output ports by restricting but not preventing free fluid flow between adjacent cavities. The device also includes anodic and cathodic compartments containing electrodes for establishing an electric field across the cavities.

Multi-channel separation device

A novel pH buffering system which is functionally stable under electrophoretic conditions employs buffering components in complementary buffer pairs. The buffering components are selected from among simple chemically defined ampholytes, weak acids and weak bases, and are paired together on the basis of their dissociation characteristics to define narrow overlapping buffering zones in which the buffering components exhibit low net electrophoretic mobility combined with desired pH buffering capacity. The narrow buffering zones cover from 0.4 to 1.25 pH units within a broad range of pH values of from about pH 3 to about pH 10. Specific buffering pH values within the narrow buffering zones are obtainable by varying the molar ratios of the two buffering components of the selected buffer pair. The buffering system may be used to form functionally stable precast narrow pH zone gradients in free solution, and has particular utility in isoelectric focusing and other electrophoretic processes.

Buffering system and its use in electrophoretic processes

Chemical reactions are readily carried out using supercritical carbon dioxide as the reaction medium. Supercritical carbon dioxide is of special value as a reaction medium in reactions for synthesizing polypeptides, for sequencing polypeptides, or for amino acid analysis.

Organic syntheses employing supercritical carbon dioxide as a reaction

The disclosure is directed to a process and apparatus for isoelectric focusing and isotachophoresis of fluids. Recycling of the process fluid is established in a first direction through an electrophoretic chamber having parallel walls, the electric field being applied in a second direction. Externally of the chamber, the flow of fluid is subdivided into a plurality of closed loops, cooled to dissipate the Joule heat. While maintaining conditions of laminar flow within the chamber, unusual fluid stabilization against convective, electroosmotic and other fluid flow disturbances is obtained if the gap between the parallel walls of the chamber is kept narrow and the fluid flow is rapid, to maximize shear stress at the walls of the chamber and minimize the fluid residence time in the chamber.

Process and apparatus for recycling isoelectric focusing and

This disclosure is directed to an isoelectric focusing apparatus, wherein stabilization of the fluid containing the isolated proteins is achieved by carrying out the separation in a rotating cylinder with the separation cavity of the cylinder being segmented by means of filter elements. The filter elements are constituted of a material offering some degree of resistance to fluid convection, but allowing relatively free and unhindered passage of current and transport of proteins. The combined effect of segmentation and rotation has been found to be superior to either segmentation or rotation alone in maintaining the stability of the migrated fractions.

Rotating apparatus for isoelectric focusing

Total or fractional precipitation of proteins from salt-containing proteinaceous solution or suspension mixtures is induced or enhanced by subjecting the proteinaceous mixture to a desalting treatment, for example, by electrodialysis. In one aspect of the invention, the desalting treatment is used in conjunction with the addition of heavy metal ions to the proteinaceous mixture to provide a synergistic effect on the precipitation of the protein fraction. In another aspect of the invention, the desalting treatment, either with or without the heavy metal ion addition, is employed as an integral part of a selective plasmapheresis procedure for the removal or recovery of one or more selected plasma proteins from blood plasma.

Precipitation of proteins from salt-containing proteinaceous fluids

The present invention relates to the reduction of the fibrinogen concentration in cryoprecipitate fraction formed during the fractionation of blood. This zinc-mediated decrease of fibrinogen increases the ratio of Factor VIII to total protein content.

Purification of antihemophilia factor VIII by precipitation with zinc

The disclosure is directed to an apparatus for isoelectric focusing. A fluid is employed which contains buffering compounds capable of establishing a pH gradient in an electric field. A plurality of ion non-selective permeable membranes are used to streamline the flow of fluid while allowing interchange of fluid constituents therebetween. Electrodes establish an electrical potential transverse the flow of fluid to establish a gradient of pH steps as between successive channels defined by the membranes.

Isoelectric focusing apparatus

The disclosure is directed to a method and apparatus for isoelectric focusing of fluids. In accordance with the disclosed method, the flow of fluids to be processed is established in a first direction. This flow of fluids is streamlined by providing a plurality of permeable microporous membranes which define generally parallel channels oriented in the first direction. An electrical potential is applied across the streamlined channels of flowing fluid, and isoelectric focusing is achieved on the fluids during the flow thereof since the membranes allow interchange of fluid constituents therebetween while providing the desired streamlining. An approximation of 'plug' type flow is achieved within the streamlined channels; i.e., flow having an approximately uniform cross-sectional characteristic. In the preferred embodiment of the method of the invention, a recirculation path is established for each of the streamlined channels, such that the fluid flowing out of each channel is recirculated back to the beginning of the channel. Preferably, this is achieved by pumping the fluids in each of the recirculation paths, and also providing cooling for the fluids during the recirculation thereof. In this manner, a number of 'passes' are effected to obtain the desired degree of isoelectric focusing. The cooling, which is performed during the recirculation, serves to minimize problems with dissipation of Joule heat during the isoelectric focusing of the fluids in the streamlined channels. Also, in the preferred embodiment of the invention, the first direction is downward such that the streamlined fluids flow under the influence of gravity to permit gravity equilibrium of fluid levels in the channels across the streamlining membranes.

Isoelectric focusing method

Electrophoretic fractional elution apparatus has a column with a rotating seal joint at which a thin jet of eluting buffer is directed across the lumen of the electrophoretic column in a direction perpendicular to that of electrophoretic migration. Either the content of the column is rotated with respect to the stationary jet, or the jet is rotated with respect to the column. The system may employ electrophoresis either in free solution or in packed columns.

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Tucson, AZ, United States of America

Milan Bier