Abstract: Methods for profiling oligosaccharides released from glycoproteins and methods for the electrophoretic separation of high mannose, complex and hybrid oligosaccharides are disclosed. The disclosed methods include first providing an analytical sample which includes at least one released oligosaccharide and introducing the sample into an electrophoretic channel containing an electrophoretic separation medium having a pH less than 8 and greater than 2.5.Then applying a sufficiently high electric field causes the released oligosaccharides to electrophoretically migrate and separate within the electrophoresis channel in accordance with their molecular size or charge to mass ratio. Detecting the separated oligosaccharides provides glycoprotein conjugate profiling information. Preferred embodiments include using an electrophoresis separation medium which includes a lithium acetate buffer having pH of about 4.75 and a gel of about 0.4 wt % polyethylene oxide.

Abstract: A gel pump operated by a stepper motor may be used to deliver fresh gel to one or more capillaries through one or more manifolds and valves. By controlling the valves, fresh gel delivered by the pump will replace the old gel in the capillaries for an automated gel replacement system. In a different setting of the valves, the manifolds may be purged of the old gel prior to delivery of fresh gel to the capillaries. The gel delivery system may also be combined with an electrophoresis system so that sealing connection capable of withstanding high pressure adequate for gel injection need not be frequently broken when the gel is to be replaced. Manifold/reflector assembly is advantageously used in the system that facilitates electrical circuit for electrophoresis and for gel replacement.

Abstract: Coated capillary electrophoresis columns and methods for their use in electrophoretic separations are disclosed. The coated capillary columns include a length of tubing having an interior surface having an interconnected polymeric coating. The interconnected polymeric coating includes a hydrophobic hydrocarbon polymeric functionality covalently bound to the interior surface and polyvinylalcohol interconnected with the hydrophobic polymeric functionality. Exemplary columns are prepared by causing a Si-OH reactive compound, having a hydrophobic polymeric functionality, to react with Si-OH functionalities on the interior surface of capillary columns. Then causing vinyl acetate to polymerize in contact with the hydrophobic polymeric functionality and hydrolyzing the resulting polyvinylacetate forms a coating of interconnecting hydrophilic and hydrophobic polymers.

Abstract: A Capillary Electrophoresis apparatus and method are disclosed which utilize a meltable plug in an end of the capillary tube to selectively pass small ionic contaminants electrophoretically and retain macromolecular analytes against one end of the plug until the plug is melted. When this plug is melted, the analytes and the contaminants pass through unimpeded. This permits separation of the analytes from the contaminants during electrophoretic separation and enhances instrument resolution.

Abstract: The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification ("base calling") is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations.

Abstract: Separation media for electrophoresis, and methods of filling and flushing of electrophoretic devices such as capillaries are described. By preparing submicron to above-micron sized cross-linked gel particles and using gel swelling equilibrium concepts, such devices can be easily filled and flushed. Gel particles can be prepared by inverse suspension, precipitation and suspension polymerization. These particles can be swollen and collapsed by small changes in temperature, pH, and ionic strength of solvent. Other approaches involve the formation of reversible cross-links by use of polyelectrolyte complexes, chelating agents or copolymers of hydrophobic and hydrophilic repeat units. Finally, reversibly solubilized systems may be used to change the viscosity of the media.

Abstract: The fraction collector according to the present invention comprises a separating means composed of electrophoresis tracks to separate samples by electrophoresis, and a transferring means to transfer the separated components eluted from said electrophoresis tracks, a transferring means which comprises the capillary sample transferring tubes which are placed with their ends close to the ends of said electrophoresis tracks at the specified gap, and which transfers the separated components eluted from each electrophoresis track, a transport means to supply the buffer solution to said gap and to carry said separated component to said sample transferring tube by sheathflow of said buffer solution, a fractionating means to fractionate the separated components having moved inside said sample transfer tube, a detecting means to detect the elution of said separated component by detecting the light emitted from said separated component, and a control means to control said fractionating means based on the signal gained b

Abstract: A capillary electrophoresis separation matrix for single-stranded nucleic acids, along with methods for using and preparing the matrix, are disclosed. The separation matrix provides denaturing conditions and contains hydroxyethyl cellulose (HEC) in combination with urea, and preferably also includes formamide. The separation matrix may be used for DNA sizing and sequencing applications and provides a single-base resolution to approximately 500 base pairs. The separation matrix is inexpensive, easy to prepare, requires no polymerization steps, and is of low enough viscosity to be pumped easily into and out of capillary tubes for electrophoresis. The low viscosity allows for high throughput of samples and reuse of the capillary tubes for numerous separations.

Abstract: Described is a method of preparing polyacrylamide gel containing capillary tube wherein acrylamide is polymerized from a liquid to a gel within the tube, the improvement comprising the steps of: a) after the polymerization, annealing the gel inside the tube by cycling the gel/tube at least once in a temperature range from 20.degree. C. to a temperature not to exceed about 75.degree. C. and back to ambient; and b) recovering the stress released gel-filled tube. Preferably, the polymerization commences at the middle of the tube and commences toward the extremities of the tube.

Abstract: An electrophoresis apparatus for detecting samples migrating in migration portions includes a plurality of gel separation portions separate from one another and having respective ends disposed along a straight line, a plurality of migration portions equal in number to the gel separation portions and disposed along a straight line following respective ones of the gel separation portions, a light source for generating light which passes through the migration portions, the light propagating along a straight line extending through all of the migration portions, a plurality of optical fibers having respective first ends and respective second ends, the first ends of the optical fibers being disposed facing respective points where the light from the light source passes through respective ones of the migration portions, the optical fibers being equal in number to or greater in number than the migration portions, and an optical detector optically coupled to the second ends of the optical fibers for receiving light fro

Abstract: A capillary electrophoresis method and apparatus for reducing dispersion of sample fractions are disclosed. The capillary tube in which the electrophoresis is performed has been flared, at least at its sample entrance end, to remove sharp corners which contribute to aberrations in the electric field distribution in a radial direction and result in differential migration of molecules depending on their proximity to the sharp corners. The flared tube, in contrast to a conventional tube, provides a more uniform electric field for electrophoresis and reduces undesired dispersion of the samples.

Abstract: The invention relates to a process for deactivating the inner surfaces of capillaries for capillary zone electrophoresis and capillary gel electrophoresis. The deactivation is effected by coating the inner surface with polar polymers which initially are water-soluble, whereupon the polymers are subsequently thermally fixed by a formation of water-insoluble polymers. The invention further relates to the capillaries thus produced and to the use thereof for the separation of oligonucleotides, peptides and proteins.