Chemistry: electrical and wave energy – Processes and products – Electrostatic field or electrical discharge
Patent
1989-02-02
1991-02-19
Niebling, John F.
Chemistry: electrical and wave energy
Processes and products
Electrostatic field or electrical discharge
204301, 2041823, 2041828, 204299R, G01N 2726, G01N 2728
Patent
active
049941615
ABSTRACT:
A method of (and apparatus for) determining charge on protein or other macromolecule. The method involves providing two spaces, each containing buffer (e.g., water or saltwater), establishing a volume (a cuvette herein) between the two spaces, which volume contains the macromolecule in a solution that includes the buffer that establishes the pH of the solution (typically a pH of 0 to 12) and that further includes a gelling agent to inhibit convection. The macromolecule is confined to the volume (the cuvette); there is electrical continuity through the volume from the buffer in one space to the buffer in the other space; and there is flow of small molecules (e.g., of the buffer) from one space to the other through the volume. There is also external flow of buffer between the two spaces to maintain constancy of ionic contact therein as well as constancy and equality of liquid level, but no electric current flow between the spaces through or along the buffer in the external flow. A d-c electric field is established across the volume from one space to the other to establish an electrophoretic force on each macromolecule in the volume to cause each macromolecule to migrate therein, the diffusive force within the volume tending to counteract the electrophoretic force. The diffusive force is opposite to the electrophoretic force. After equilibrium between the two forces has occurred, the concentration distribution of macromolecule is determined and the slope of the log of the concentration as a function of protein or curve fitting directly to the concentration direction is used to arrive at the charge on the macromolecule.
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Hazard Andrea L.
Laue Thomas M.
Yphantis David A.
Niebling John F.
Phasge Arun S,.
Shaw Robert
University of New Hampshire
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