Biomolecule Stability Measurement Using Localized Laser Heating
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Solution Overview
Problem
Conventional methods for measuring biomolecule stability and interactions are time-consuming, often taking over an hour, and lack sensitivity to surface properties and conformational changes due to their reliance on size changes and thermal mass equilibration.
Innovation Solution
A method utilizing infrared laser radiation to create localized, high-temperature gradients for rapid thermo-optical characterization of biomolecules, allowing for contact-free measurements within 1 ms to 200 ms, enabling the detection of thermophoretic mobility and hydrodynamic radius, and discrimination between different conformations and species.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional homogeneous heating methods are used to measure biomolecule stability, then the entire sample volume reaches the applied temperature, but the measurement process becomes very time-consuming (over an hour) due to the large thermal mass requiring long equilibration times
Solution Approach 1:
The patent applies local quality by creating highly localized temperature gradients using focused laser beams that heat only specific regions of the sample containing biomolecules. This localized heating approach allows rapid temperature changes in the measurement region without requiring the entire sample volume to equilibrate, thereby achieving fast biomolecule stability measurements (reducing time from over an hour to seconds) while maintaining measurement precision through targeted thermal exposure of the biomolecules of interest
2Stability of the object's composition
If stepwise temperature increase with waiting time is used for each temperature point, then the system reaches thermal equilibrium, but the overall heating process takes a very long time
Solution Approach 1:
The patent employs periodic action by using pulsed laser heating to create rapid, cyclic temperature changes in the sample. Instead of continuous stepwise heating with long equilibration periods, the system applies short laser pulses that periodically heat the biomolecules, allowing thermal equilibrium to be achieved rapidly in the measurement region while maintaining high productivity through efficient energy delivery and rapid thermal response
3Reliability
If contact-free optical methods are used for biomolecule characterization, then sample contamination is avoided, but the measurement technique lacks sensitivity to surface properties and conformational changes
Solution Approach 1:
The patent applies parameter changes by utilizing temperature as a controlled parameter to induce and detect conformational changes in biomolecules through their intrinsic fluorescence properties. By systematically varying temperature and monitoring fluorescence intensity changes, the method achieves sensitivity to surface properties and conformational states while maintaining contact-free measurement reliability, as the thermal energy modifies molecular parameters that directly affect fluorescent emission characteristics
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
Enables fast and sensitive analysis of biomolecular interactions and stability, reducing measurement time significantly compared to existing methods, and providing insights into surface properties and conformational changes.
Implementation Method 1
irradiating a laser light beam into the solution to obtain a spatial temperature distribution in the solution around the irradiated laser light beam
Implementation Method 2
The temperatures are measured with high spatial resolution by the temperature sensitive fluorescence of a fluorescent dye
Data Source
Figure 1a~1b
Figure 2
Figure 3a~3d
AI summary
The present invention relates to a method for measuring the stability, for example based on the detected unfolding, of a biomolecule, such as a protein, peptide, antibody or nucleic acid, in a solution comprising: (a) providing a sample with the biomolecule in a solution; (b) exciting fluorescently the biomolecule and detecting a first fluorescence of the excited biomolecule; (c) irradiating a laser light beam, preferably a focused infrared laser light beam, into the solution to obtain a spatial temperature distribution in the solution around the irradiated laser light beam; (d) detecting a second fluorescence of the biomolecule in the solution at a predetermined time after irradiation of the laser into the solution has been started, and (e) characterizing the stability of the biomolecule based on the first and second detections.