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Ratiometric Two-Photon Near-Infrared Probe to Detect DPP IV in Human Plasma, Living Cells, Human Tissues, and Whole
Javier Valverde-Pozo1, Jose M Paredes1, Thomas J Widmann2
1Nanoscopy-UGR Laboratory, Department of Physical Chemistry, Faculty of Pharmacy, Unidad de Excelencia en Quimica Aplicada a Biomedicina y Medioambiente (UEQ), University of Granada, C. U. Cartuja, 18071 Granada, Spain.
ACS Sensors
|February 27, 2023
Summary
Researchers developed a novel near-infrared (NIR) probe to detect dipeptidyl peptidase IV (DPP IV) activity. This probe enables rapid, efficient detection in living cells, tissues, and organisms, overcoming limitations of traditional methods.
Area of Science:
- Biochemistry
- Molecular Biology
- Medical Diagnostics
Background:
- Dipeptidyl peptidase IV (DPP IV), also known as CD26, is a transmembrane glycoprotein involved in glucose metabolism and T-cell activation.
- DPP IV is overexpressed in several human carcinomas and can serve as a diagnostic marker for lysosomal storage diseases.
- Accurate detection of DPP IV activity is crucial for understanding physiological and disease states.
Purpose of the Study:
- To design and synthesize a novel near-infrared (NIR), ratiometric, two-photon (TP) fluorimetric probe for DPP IV activity.
- To develop a sensitive and efficient method for detecting DPP IV in various biological samples.
Main Methods:
- A ratiometric NIR fluorimetric probe was constructed by conjugating a Gly-Pro recognition group to a two-photon (TP) fluorophore (DCM-NH2 derivative).
- The probe's fluorescence properties were characterized, focusing on the restoration of its internal charge transfer (ICT) emission spectrum upon enzymatic cleavage.
- DPP IV activity was detected in living cells, human tissues, and zebrafish using the developed probe and TP excitation.
Main Results:
- The designed probe exhibits high ratiometric fluorescence output upon specific enzymatic release of the dipeptide group.
- The probe enabled rapid and efficient detection of DPP IV activity in living cells, human tissues, and zebrafish.
- Two-photon excitation allowed for the detection of DPP IV activity in plasma, circumventing autofluorescence and photobleaching issues associated with visible light excitation.
Conclusions:
- The novel NIR, ratiometric, TP fluorimetric probe provides a sensitive and efficient tool for detecting DPP IV enzymatic activity.
- This probe facilitates non-invasive imaging and diagnostics in various biological systems, including living organisms.
- The probe's ability to avoid autofluorescence and photobleaching enhances its utility for detecting DPP IV activity in complex biological matrices like plasma.
