A Multichannel Time-Tagged Time-Resolved (TTTR) Model for Quantification of Oligomer Concentrations Based on...

Area of Science:

• Biophysics
• Biochemistry
• Analytical Chemistry

Background:

• Molecule and protein aggregation are implicated in numerous incurable human diseases.
• Early-stage protein oligomers are particularly toxic and contribute significantly to disease pathogenesis.
• Differentiating and quantifying various protein oligomers remains a significant challenge in fundamental research.

Purpose of the Study:

• To develop a novel method for differentiating and quantifying protein oligomers.
• To address the challenge of analyzing toxic protein aggregates in disease research.
• To establish a reliable analytical relationship between photon emission and oligomer concentration.

Main Methods:

• Development of a multichannel time-tagged time-resolved (TTTR) confocal fluorescence model.
• Utilizing the antibunching effect, where n-oligomers labeled with n-dyes emit a maximum of n photons.
• Derivation of an analytical relationship between photon emission probability and oligomer concentrations, assuming ideal dye behavior.

Main Results:

• The developed TTTR confocal fluorescence model successfully differentiates protein oligomers.
• Quantification of up to tetraoligomers was achieved with a relative error of less than 10%.
• Simulations validated the model's efficacy in an eight-channel setup with eight single-photon detectors.

Conclusions:

• The novel TTTR confocal fluorescence model offers a viable solution for protein oligomer differentiation and quantification.
• This method has significant implications for fundamental research into aggregation-related diseases.
• The model provides a foundation for improved diagnostics and therapeutic strategies targeting protein aggregation.