To develop new therapies, diagnose diseases, or understand fundamental biological mechanisms, scientists need the ability to monitor biomolecules inside living cells with precision. However, current techniques for intracellular biomolecule labeling are either imprecise, inefficient, or toxic.
One of the most powerful tools in chemical biology, copper-catalyzed azide-alkyne cycloaddition (CuAAC)—commonly known as Click Chemistry—has long been used for biomolecule labeling. Despite its widespread application, Click Chemistry has never been viable for live-cell research because it requires toxic levels of copper for cells.
Now, Dr. Sara H. Rouhanifard, Assistant Professor of Bioengineering at Northeastern University, is breaking this long-standing barrier. Her lab has developed InCu-Click, a breakthrough reagent that makes Click Chemistry safe for live-cell applications by reducing copper toxicity while enabling real-time biomolecule labeling.
With the support of the Fall 2024 Spark Fund Award, Dr. Rouhanifard’s team is expanding the potential of InCu-Click—advancing research applications, optimizing its performance, and preparing for commercialization. This innovation has the potential to transform drug discovery, biomarker detection, and fundamental cellular biology.
InCu-Click: Removing a Barrier to Studying Live Cells
For decades, researchers have been eager to track biomolecules inside living cells in real-time. However, existing intracellular labeling techniques are limited.
Without a safe, efficient way to track molecular interactions in live cells, researchers are forced to rely on indirect methods that often lack precision, require cell fixation (which kills the cells), or disrupt cellular function.
By developing an ultrasensitive copper-chelating ligand, InCu-Click allows biomolecules to be labeled inside living cells without harming them. It binds copper tightly, keeping its concentration at safe levels while still enabling the reaction to take place efficiently. Additionally, the system’s advanced delivery mechanism ensures precision in targeting biomolecules inside cells without interfering with normal cellular function.
“As soon as people are able to track things inside of live cells, it opens up entirely new research avenues,” says Dr. Rouhanifard. “It allows you to track drugs in real-time while they perform their functions inside of cells, revealing mechanisms of action, drug targets, and fundamental biological interactions.”
With this advancement, scientists will be able to observe biomolecule interactions as they naturally occur, leading to deeper insights into cell signaling, disease progression, and therapeutic interventions.
Looking Ahead: Bringing InCu-Click to the Scientific Community
InCu-Click is not just a breakthrough for academic research—it has the potential to become a standard tool in biotech and pharmaceutical labs worldwide.
The demand for live-cell biomolecule labeling technologies is rapidly increasing, particularly in fields such as molecular biology, drug discovery, and diagnostics. The global cellular imaging and labeling market is projected to exceed $15 billion by 2027, driven by expanding investments in life sciences, cell-based research, and precision medicine. With more than one million research labs worldwide working in academia, biotech, and pharmaceuticals, the adoption of InCu-Click could be transformative.
Dr. Rouhanifard envisions a future where InCu-Click is as accessible as other click-chemistry reagents, which are widely used in labs today.
“Much like the current commercial Click-IT kits are now, I hope InCu-Click becomes one of those bread-and-butter tools that every lab is using,” she says.
The ability to monitor molecular interactions inside living cells could accelerate the development of targeted therapies, improve diagnostics, and unlock new frontiers in biological research.
The Spark Fund: Driving Innovation from Lab to Market
For this work, Dr. Rouhanifard received the Fall 2024 Spark Fund Award from Northeastern University.
The Spark Fund will allow her team to push InCu-Click beyond the research lab and into broader applications. They plan to expand their research team, optimize the reagent for broader applications, including live-animal research, and explore commercialization strategies to bring InCu-Click to the global scientific community.
“The Spark Fund process helped us refine how we think about the real-world applications of our research,” she explains. “Industry experts asked us questions we had never considered, and it helped push our thinking beyond the academic lab.”
With continued research and strategic partnerships, the team hopes that InCu-Click could soon become an indispensable tool in life sciences, accelerating discoveries in drug development, disease research, and molecular biology.
“We’ll keep pushing the limits of this and seeing if we can broaden the utility in the lab and for more use cases,” she says.
Key Takeaways: A New Era for Live-Cell Research
InCu-Click represents a major leap forward in biomolecule labeling, providing a safe, efficient, and precise method to track molecular interactions inside live cells. This breakthrough removes long-standing barriers to live-cell research, opening up new possibilities in drug discovery, biomarker development, and disease modeling.
With the support of Northeastern’s Spark Fund, Dr. Rouhanifard and her team are advancing this technology to become a vital tool for the next generation of scientific discovery, unlocking the way for groundbreaking advancements in medicine and biotechnology.
Learn More about the Spark Fund
Learn more about Rouhanifard’s research and the three other Fall 2024 Spark Award awardees here.
Written by Elizabeth Creason