Northeastern University

Massachusetts Institute of Technology

Beckman Coulter, Inc.

Frantisek Foret

Barry L Karger

Lev Kotler

Marek Minarik

Karel Kleparnik

Jan Preisler

Paul M Zavracky

Roger W Giese

William G Thilly

Aharon S Cohen

Yuriy M Dunayevskiy

Thomas Rejtar

Bailin Zhang

Phouthone Koehavong

Konstaintin Khrapko

Qifeng Xue

E Nicol McGruer

Daniel P Kirby

Dieter Schmalzing

Odilo M Uller

A modular multiple lane or capillary electrophoresis (chromatography) system that permits automated parallel separation and comprehensive collection of all fractions from samples in all lanes or columns, with the option of further on-line automated sample fraction analysis, is disclosed. Preferably, fractions are collected in a multi-well fraction collection unit, or plate. The multi-well collection plate is preferably made of a solvent permeable gel, most preferably a hydrophilic, polymeric gel such as agarose or cross-linked polyacrylamide.

Multichannel microscale system for high throughput preparative separation with comprehensive collection and analysis

While the present invention has been described in conjunction with a preferred embodiment, one of ordinary skill, after reading the foregoing specification, will be able to effect various changes, substitutions of equivalents, and other alterations to the compositions and methods set forth herein. It is therefore intended that the protection granted by Letters Patent hereon be limited only by the definitions contained in the appended claims and equivalents thereof.

A sample load and injection device () includes sample introduction capillaries () attached to a microfluidic device (). Sample introduction capillaries () are connected to sample introduction channels (). Sample introduction capillaries () are connected to separation channels () at connection points (). At a defined distance along the separation channels (), auxilliary channels () originate at connection points (). The sample load and injection device includes cover plate () which has connecting channels (). Connecting channel () is associated with ends () of sample introduction channels (). Connecting channel () is associated with ends ().

Parallel sample loading and injection device for multichannel microfluidic devices

A thermostat array including an array of two or more capillary columns () or two or more channels in a microfabricated device is disclosed. A heat conductive material () surrounded each individual column or channel in array, each individual column or channel being thermally insulated from every other individual column or channel. One or more independently controlled heating or cooling elements () is positioned adjacent to individual columns or channels within the heat conductive material, each heating or cooling element being connected to a source of heating or cooling, and one or more independently controlled temperature sensing elements () is positioned adjacent to the individual columns or channels within the heat conductive material. Each temperature sensing element is connected to a temperature controller.

Parallel array of independent thermostats for column separations

A universal interface for continuous on-line liquid sample introduction directly to the time-of-flight mass spectrometer, which can further promote throughput and utility of MALDI-TOF MS, is disclosed. Preferably, the liquid sample includes a matrix, either solid or liquid, for use in matrix-assisted-laser-desorption-ionization, most particularly in a time-of-flight mass spectrometer which can further promote throughput and utility of MALDI-TOF MS. In the method of the invention, the same samples and matrices, both solid and liquid, can be used as in conventional MALDI. In practice of the method of the invention, a solution of sample containing, e.g., peptide and matrix is infused directly into the source chamber of a mass spectrometer at subatmospheric pressure, deposited on a moving sample holder, such as a rotating quartz wheel, and desorbed by, e.g., a nitrogen laser. The method of the invention is particularly amenable to multiplexing, the parallel deposition of multiple samples, e.g., from a capillary array or microchip channels, with subsequent sequential desorption with a scanning laser. This format is particularly useful for high throughput MS analysis. Also disclosed is an off-line deposition chamber and a general method of preparing a sample for analysis that results in the homogeneous deposition of small quantities of sample at improved reproducibility on any appropriately configured sample receptor. This format of sample preparation is particularly useful with existing commercial mass spectrometers.

On-line and off-line deposition of liquid samples for matrix assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectroscopy

A universal interface for continuous on-line liquid sample introduction directly to the time-of-flight mass spectrometer, which can further promote throughput and utility of MALDI-TOF MS, is disclosed. Preferably, the liquid sample includes a matrix, either solid or liquid, for use in matrix-assisted-laser-desorption-ionization, most particularly in a time-of-flight mass spectrometer which can further promote throughput and utility of MALDI-TOF MS. In the method of the invention, the same samples and matrices, both solid and liquid, can be used as in conventional MALDI. In practice of the method of the invention, a solution of sample containing, e.g., peptide and matrix is infused directly into the source chamber of a mass spectrometer at subatmospheric pressure, deposited on a moving sample holder, such as a rotating quartz wheel, and desorbed by, e.g., a nitrogen laser. The method of the invention is particularly amenable to multiplexing, the parallel deposition of multiple samples, e.g., from a capillary array or microchip channels, with subsequent sequential desorption with a scanning laser. This format is particularly useful for high throughput MS analysis. Also disclosed is an off-line deposition chamber and a general method of preparing a sample for analysis that results in the homogeneous deposition of small quantities of sample at improved reproducibility. This format of sample preparation is particularly useful with existing commercial mass spectrometers.

A modular multiple lane or capillary electrophoresis (chromatography) system that permits automated parallel separation and comprehensive collection of all fractions from samples in all lanes or columns, with the option of further on-line automated sample fraction analysis, is disclosed. Preferably, fractions are collected in a multi-well fraction collection unit, or plate ( ). The multi-well collection plate ( ) is preferably made of a solvent permeable gel, most preferably a hydrophilic, polymeric gel such as agarose or cross-linked polyacrylamide.

A universal interface for continuous on-line liquid sample introduction directly to the time-of-flight mass spectrometer, which can further promote throughput and utility of MALDI-TOF MS, is disclosed. Preferably, the liquid sample includes a matrix, either solid or liquid, for use in matrix-assisted-laser-desorption-ionization, most particularly in a time-of-flight mass spectrometer which can further promote throughput and utility of MALDI-TOF MS. In the method of the invention, the same samples and matrices, both solid and liquid, can be used as in conventional MALDI. In practice of the method of the invention, a solution of sample containing, e.g., peptide and matrix is infused directly into the source chamber of a mass spectrometer at subatmospheric pressure, deposited on a moving sample holder, such as a rotating quartz wheel, and desorbed by, e.g., a nitrogen laser. The method of the invention is particularly amenable to multiplexing, the parallel deposition of multiple samples, e.g., from a capillary array or microchip channels, with subsequent sequential desorption with a scanning laser. This format is particularly useful for high throughput MS analysis.

On-line liquid sample deposition interface for matrix assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectroscopy

A subatmospheric, variable pressure sample delivery chamber, useful as an electrospray ionization device for introducing a sample into a mass spectrometer or as a sample delivery device for delivery of a sample to a collection device at subatmospheric pressures, is disclosed. The sample delivery chamber is configured to maintain an operating pressure between 1 and 750 Torr, preferably between 50 and 700 Torr. The chamber has an inlet port for introduction of a gas and an exit port capable of being coupled either to a sampling orifice for an analytical device, usually a mass spectrometer, or directly to a pump for removal of gas. A sample delivery device extends from outside the chamber into the interior of the chamber. The inlet end of the sample delivery device is capable of receiving a sample and the exit end of the sample delivery device is aligned with the exit port of the chamber, if the sample is to be delivered to an external analytical device, or aligned with the collection device, for direct collection of the sample.

Subatmospheric, variable pressure sample delivery chamber for

A microscale fluid handling system that permits the efficient transfer of nanoliter to picoliter quantities of a fluid sample from the spatially concentrated environment of a microfabricated chip to 'off-chip' analytical or collection devices for further off-chip sample manipulation and analysis is disclosed. The fluid handling system is fabricated in the form of one or more channels, in any suitable format, provided in a microchip body or substrate of silica, polymer or other suitable non-conductive material, or of stainless steel, noble metal, silicon or other suitable conductive or semi-conductive material. The microchip fluid handling system includes one or more exit ports integral with the end of one or more of the channels for consecutive or simultaneous off-chip analysis or collection of the sample. The exit port or ports may be configured, for example, as an electrospray interface for transfer of a fluid sample to a mass spectrometer.

Microscale fluid handling system

The disclosure relates to a method for resolving double-stranded DNA species differing by at least one base pair. Each of the species is characterized by an iso-melting domain with a unique melting temperature contiguous with a melting domain of higher thermal stability.

Electrophoretic detection and separation of mutant DNA using replaceable

A capillary electrophoretic system and method for accurate and precise post-column manipulation of molecules separated by capillary electrophoresis is disclosed. The system of the invention includes a separation capillary; a detector positioned close to, and preferably less than approximately one cm from the outlet end of the capillary; and a sheath which surrounds and directs a collection buffer over the capillary outlet end. With the detector positioned as described, precise and accurate correlation is possible between detection of the separated components of a sample and their emergence from the capillary. The detector preferably includes optical fibers connected to a spectrophotometric detection method, such as UV detection or fluorescence, including laser induced fluorescence. The sheath collection buffer, the capillary and an electrophoresis buffer reservoir, for supplying buffer to the inlet end of the capillary, are in electrical contact in an electrical circuit and provide the electric field required for separation of a sample of molecules. The separated components of the sample are mixed with collection buffer as they emerge from the outlet end of the capillary. They are then collected into or on any collection device, such as vials, collection capillaries or membranes, or they are transferred to other devices for further analysis, such as mass spectroscopy.

Precise capillary electrophoretic interface for sample collection or

A coated microcapillary column for high performance electrophoresis is disclosed. A preferred microcapillary includes a column; a universal sub-layer of coating material that has been highly crosslinked using a crosslinking agent and attached covalently or non-covalently to the column wall; and a variable top layer of a monomer or polymer, preferably crosslinked, attached to the sub-layer. The microcapillary preferably is prepared by covalently bonding a highly crosslinked siloxanediol sub-layer to the inner surface of the microcapillary wall and then causing a mixture of monomers or polymers to react in the bore of the microcapillary to form a top layer. The bilayer coating as used in a microcapillary prevents adsorption of solutes during electrophoresis and eliminates or controls electroosmotic flow. The disclosed coating may also be formed on other kinds of surfaces where similar surface modification is desired. The sub-layer, which can be formed from other siloxane derivatives having two nucleophilic end groups, e.q., diamino or dithiol, may be used independently, or as a base layer for bilayer or even multilayer surface formation.

Growing community of inventors

Malden, MA, United States of America

Frantisek Foret