Apparatus for separating molecules

Chemistry: electrical and wave energy – Processes and products – Electrophoresis or electro-osmosis processes and electrolyte...

Reexamination Certificate

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C204S450000, C204S456000, C204S466000, C204S600000, C204S606000, C204S616000, C204S643000

Reexamination Certificate

active

06296752

ABSTRACT:

The present invention relates to methods of and apparatuses for separating molecules, such as proteins or nucleic acids. The invention further relates to apparatuses that allow, and methods that use, an integrated mass spectrometric analysis.
SUMMARY OF THE INVENTION
Provided is an apparatus for separating, in a medium, a component from a composition comprising:
(1) an array of three or more electrodes arrayed along a pathway along which molecules of the composition are transported; and
(2) a power source device for delivering to voltage to the electrodes; wherein the voltages delivered to the electrode array by the power source device are effective to:
(a) alter the relative movement along the transport pathway of two or more of the molecules caused by a motive force, or
(b) cause the molecules to move along the transport pathway.
The apparatus can further comprise: (3) where the electrode array and power source device do not provide a primary motive force for moving the molecules along the transport pathway, a source of motive force comprising (i) a source of pressure for promoting bulk fluid flow along the transport pathway or (ii) electrodes for promoting electrophoretic transport of the molecules along the transport pathway. In one embodiment, the three or more electrodes are separated from the fluid by a dielectric sufficient to prevent charge extraction or insertion from the electrodes to the fluid. In another embodiment, the three or more electrodes contact the fluid.
In an embodiment, the power source is programmed to deliver voltage to each of three or more said electrodes by periodically reversing the voltage polarity applied to the electrodes with a frequency of at least 10 Hz while maintaining a desired net effect. Preferably, the power source is programmed to deliver voltage to each of three or more said electrodes over an operating period of time encompassing at least one polarity cycle satisfying either (a) a first ratio of a voltage-integrated area A
1
associated with a first polarity to a voltage-integrated area A
2
associated with the other polarity or (b) a second ratio of a charge q
1
carried by the current associated with a first polarity to a charge q
2
carried by the current associated with the other polarity is less than 1: ½ and more than about ½:1.
In an embodiment, the apparatus has the three or more electrodes are located along one face of transport pathway, and the apparatus further comprises at least one counter electrode on another face of the transport pathway.
In an embodiment, the power source is programmed for delivering a traveling wave pattern of voltages to the electrodes, which traveling wave pattern of voltages is, in some embodiments, effective to move molecules in the fluid medium.
The invention provides a separation device comprising:
(1) an array of three or more first electrodes arrayed along a first pathway along which the molecules are transported; and
(2) an array of three or more second electrodes arrayed along a second pathway, which diverges from the first pathway, along which the molecules are transported,
wherein the first and second electrodes are effective to provide a motive force for moving molecules along the first or second pathway or for modifying the transport properties of molecules traveling along the first or second pathway.
The invention further provides a separation and analysis device comprising:
(1) a chamber formed between a first support and a second support, the chamber defining a region in which a separation medium can be placed, wherein the first support and second support can be separated;
(2) an array of three or more electrodes arrayed along a pathway found in the chamber, along which pathway the molecules are transported;
(3) a power source device for delivering to voltage patterns to the electrodes; and
(4) a source of focused ionizing radiation adapted to be directed to a chamber surface of the first or second support when the first and second supports are separated.
Also provided is a method of separating a component from composition in an apparatus according to claim
1
, the method comprising: moving the molecules under the power of the voltage applied to the electrodes. In one embodiment, the method comprises applying a traveling wave voltage pattern to the electrodes to move the molecules. In another embodiment the method comprises applying a motive force separate from said voltage driven electrodes to move the molecules; and altering the mobility of at least one component by applying voltage to said electrodes.
Further provided is a method of separating a component from composition and analyzing the component, the method comprising: separating the component by operating an apparatus of the invention; directing a source of ionizing radiation at the component in situ in the medium to generate ions from the component; and analyzing the component by mass spectroscopy.
Also provided is a method of separating a component from composition and analyzing the component, the method comprising: separating the component by operating an apparatus of the invention, wherein the apparatus is operated with a membrane adjacent to the counter-electrode(s) and to the medium; transferring in situ, under the influence of an electric field, the component to the membrane by operating the electrodes in conjunction with the counter-electrode(s); directing a source of ionizing radiation at the component in the membrane to generate ions from the component; and analyzing the component by mass spectroscopy.


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