Back to results
US 12,016,688 B2Granted
In vivo radical dosimetry and in vivo hydroxyl radical protein foot-printing
AssigneeGennext Technologies, Inc.
InventorScot Randy Weinberger
Life of this patent
Estimating the term…
Abstract
Systems and methods of in vivo hydroxyl radical protein foot-printing are presented. These teachings may be used to, for example, study three-dimensional protein structure or bio-kinetics. Radical Dosimetry including an optional intrinsic standard is used on isolated intact cells. Real-time feedback based on an internal standard provides comparability between different experiments and in vivo analysis results in data that is representative of actual biological conditions.
Claims30
An analysis system comprising:
- a sample introduction system configured to provide intact biological entities to a photolysis zone, the biological entities being isolated from each other in a focused sheath flow;
- a photolysis light source configured to generate light to generate hydroxide radicals from a source of hydroxide radicals;
- a photolysis zone configured to receive the sheath flow and the light so as to oxidize an internal standard and so as to oxidize biological compounds of the biological entities in vivo;
- a dosimetry zone configured to receive the biological entities from the photolysis zone, and to detect oxidation of the internal standard using a fluorescence detector;
- control logic configured to determine that a target concentration of hydroxide radicals was generated for each of the biological entities and to adjust operation of the photolysis zone to meet the target concentration of hydroxide radicals;
- and
- a reservoir configured to receive the biological entities including the oxidized biological compounds.
4 more elements
A method of oxidizing biomolecules within an intact cell, the method comprising:
- introducing a sample mixture containing at least one cell into a photolysis zone, a source of hydroxide radicals and a dosimeter internal standard into a photolysis zone of a flash photolysis system;
- providing light to generate the hydroxide radicals from the source of hydroxide radicals, the hydroxide radicals being configured to oxidize biomolecules within the at least one cell;
- waiting a time for the at least one cell to reach a dosimetry zone of a radical dosimeter configured to detect a photometric property of the dosimeter internal standard resulting from reaction of the dosimeter internal standard and the hydroxide radicals, wherein the at least one cell is detectable within a dosimetry zone of the radical dosimeter by light scattering;
- measuring a photometric property of the dosimeter internal standard using the radical dosimeter, while the at least once cell is within the dosimetry zone;
- determining that a target level of hydroxide radicals was not generated based on the measured photometric property of the dosimeter internal standard;
- and
- adjusting a concentration of hydroxide radicals within the photolysis zone by adjusting at least one of:
- 1) an amount of light provided to the photolysis zone, 2) a concentration of the source of hydroxide radicals, 3) a flow rate of the at least once cell within the photolysis zone, or 4) adjusting a time of providing the light to the photolysis zone.
5 more elements
Dependencies are inferred from the claim text.