Label-Free Evaluation of Advanced Glycation End Products by a Comprehensive Biophysical Analysis
Ameera K1, Darshan Chikkanayakanahalli Mukunda1,2, Subhash Chandra1
1Department of Biophysics, Manipal School of Life Sciences, Manipal Academy of Higher Education, Manipal, Karnataka576104, India.
Analytical Chemistry
|July 27, 2026
Summary
A new LED-induced autofluorescence method accurately detects advanced glycation end products (AGEs) on proteins. This cost-effective technique aids in diagnosing diabetes, aging, and neurodegenerative disorders by monitoring AGEs formation.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Biomedical Engineering
Background:
- Nonenzymatic glycation forms advanced glycation end products (AGEs), linked to diabetes, neurodegenerative disorders (NDDs), and aging.
- Accurate detection of protein-bound AGEs is crucial for understanding their pathology.
- Existing AGEs detection methods are often costly, time-consuming, and require complex sample preparation, hindering accurate determination due to AGEs' heterogeneity.
Purpose of the Study:
- To develop a sensitive, precise, and cost-effective autofluorescence-based methodology for detecting and tracking AGEs formation on proteins.
- To overcome the limitations of conventional AGEs detection methods.
- To provide a tool for routine monitoring of AGEs accumulation in various diseases.
Main Methods:
- Utilized a low-power fiber-coupled near-UV LED (340 nm) and a blue LED (430 nm) for excitation, covering heterogeneous excitation maxima of AGEs.
- Employed an autofluorescence-based methodology to capture AGEs-specific signatures, distinguishing them from protein autofluorescence.
- Validated the device's sensitivity using standard pentosidine and monitored the formation of various AGEs on different glycated proteins in vitro.
Main Results:
- The LED-induced autofluorescence device demonstrated high sensitivity in detecting standard pentosidine (3.66 pg/μL) at 340 nm excitation.
- The method effectively monitored the formation and progression of clinically relevant AGEs (pentosidine, argpyrimidine, vesperlysine A, B, C) on diverse glycated proteins.
- The technique successfully captured heterogeneous AGEs signatures without interference from protein autofluorescence, revealing AGEs composition.
Conclusions:
- The developed autofluorescence methodology offers a rapid, reliable, and cost-effective approach for assessing protein glycation.
- Low-power LEDs provide stable excitation, ensuring high reproducibility for AGEs detection.
- This technique facilitates routine monitoring of AGEs accumulation, advancing research in diabetes, aging, and NDDs.
