Scientists have created an innovative method to count labeled proteins within living cells, which may become an essential and beneficial instrument in biomedical research.
Scientists have created an innovative method to count labeled proteins within living cells, which may become an essential and beneficial instrument in biomedical research.
This advanced technique, called Protein-tag Degree of Labelling (ProDOL), offers a strong and adaptable method to accurately assess labeling efficiencies—the number of proteins marked with fluorescent tags in living cells—during microscopy. This is a critical element in quantifying proteins for biological studies.
A research team from the Cardiovascular Sciences and the School of Chemistry at the University of Birmingham worked alongside several international institutions to develop this method. Their findings, which emphasize ProDOL’s potential to significantly improve the accuracy and dependability of protein labeling, were recently published.
Fluorescence microscopy has been fundamental to biomedical research, allowing scientists to identify and study proteins within cells. However, the effectiveness of these analyses largely hinges on being able to determine how many proteins are tagged with fluorescent markers—a factor known as the degree of labeling (DOL). Current methods used to measure DOL have shortcomings, often leading to inconsistent labeling efficiencies and non-specific signals.
“ProDOL aims to overcome these obstacles by offering a rapid and dependable way to quantify labeling efficiencies,” stated Professor Dirk-Peter Herten, the head researcher at Cardiovascular Sciences, University of Birmingham. “This tool facilitates the refinement of protein-tagging strategies and delivers precise measurements of protein quantities within cellular structures.”
“With its demonstrated accuracy and flexibility, ProDOL is poised to become an essential and beneficial asset in biomedical research, enabling more detailed and revealing studies of cellular activities. It’s also very adaptable, suitable for various cell types and experimental settings.”
“In the future, for instance, it could enhance our comprehension of cellular signaling pathways involved in the activation of immune cells or platelets, which are crucial in the context of various ailments, including inflammation, immune deficiencies, and several cardiovascular diseases.”
Real-World Applications and Future Directions
In this research, ProDOL was utilized to explore how the HIV-1 virus influences the response of CD4 T immune cells.
Working with Professor Oliver Fackler from the Department of Infectious Diseases and Integrative Virology at Heidelberg University Hospital, the team showcased ProDOL’s capability to accurately gauge the total and activated copy numbers by counting both the overall and active quantities of helper proteins within minuscule signaling clusters of immune cells.
Furthermore, Professor Ursula Klingmüller from the Division of Systems Biology of Signal Transduction at the German Cancer Research Center (DKFZ) aided in designing the probes for live-cell applications, thus ensuring the technique’s reliability and relevance in dynamic biological environments.
