Harnessing the Power of Multispecific Antibodies

From Academic Roots to Industry Innovation

Ridhima’s journey into the world of multispecific antibodies is marked by broad scientific expertise and a seamless transition from academic roots to industry. With a solid foundation in biochemistry—gained during her bachelor’s and master’s studies in India—she further refined her research skills during her PhD and postdoctoral work in Germany, focusing on mitochondrial protein biochemistry. Although her early research centered on understanding the structure–function relationships of membrane proteins in yeast, her rigorous scientific approach and early exposure to cell-free systems provided a strong foundation for her current role at LenioBio.

“Working with a cell-free system early on, even if only in a limited way, gave me insight into rapid protein expression—a lesson I’ve carried into my work at LenioBio,” Ridhima explains. “It was a natural transition to apply that knowledge to multispecific antibodies, where precision and speed are absolutely critical.”

The Promise and Challenges of Multispecific Antibodies

Traditional monoclonal antibodies, which bind a single antigen, are integral to our immune defense. In contrast, multispecific antibodies are recombinantly engineered to target multiple proteins simultaneously. This dual (or even triple) binding capability can, for example, engage cancer cells while recruiting immune effector cells to mount a more potent attack, or facilitate the co-localization of therapeutic targets in diseases such as hemophilia or retinal disorders.

“The key benefit is generating a stronger, more tailored therapeutic response,” Ridhima notes. “Imagine one arm of the antibody binding to a cancer cell while another simultaneously recruits an immune cell—this dual action can significantly boost the immune response.”

However, this engineering feat comes with notable challenges. One significant hurdle is ensuring correct chain pairing—whether it’s heavy chain to heavy chain or heavy chain to light chain—which is critical for therapeutic efficacy and safety. Moreover, certain formats, especially fragment-based multispecifics, can be prone to aggregation. Aggregation not only compromises the stability and functionality of the antibody but also presents challenges during downstream purification processes. “For fragment-based antibodies, their smaller size makes them susceptible to aggregation during folding, which can be a major stumbling block during production,” explains Ridhima. “Addressing these challenges is essential to ensure that every batch of therapeutic antibody meets the high standards required for clinical use.”

ALiCE®: A Cell-Free Solution to Complex Protein Production

Addressing these challenges head-on is the ALiCE® platform—a cell-free protein synthesis system derived from tobacco BY-2 cells. Unlike traditional cell-based production methods that rely on HEK or CHO cell lines (often plagued by long timelines translating into high costs), ALiCE® offers a streamlined and versatile approach.

“Using the ALiCE® platform, you can express any antibody—or different protein classes—in just 24 to 48 hours,” Ridhima explains. “This is particularly advantageous during the early screening phases, where rapid iteration and optimization are critical. Instead of generating a new cell line for every construct, the same lysate can be used to express a variety of proteins. This not only saves time but also resources.”

Beyond speed, ALiCE® has already demonstrated remarkable success in producing challenging multispecific formats. LenioBio has successfully tested:

• A knob-in-hole format bispecific antibody with over 95% correct heavy chain pairing.
• A BiTE (Bispecific T-cell Engager) at high quality and with confirmed binding activity shows robust expression and functionality.
• Several tri-specific antibodies that show robust expression despite increased complexity.

These achievements underline the platform’s versatility and ability to tackle challenges such as aggregation and complex chain pairing, which are often roadblocks in traditional systems.

Bridging the Gap to Next-Generation Therapeutics

Multispecific antibodies are carving out a significant niche in personalized medicine and targeted therapeutics. Their flexibility—ranging from full-sized IgG formats to smaller fragments like nanobodies—opens up applications across oncology, coagulation disorders, and beyond. As the market for these therapeutics grows, rapid and reliable production methods become essential.

“Our roadmap is to validate more common formats such as crossmab and common light chain designs using ALiCE®,” says Ridhima. “By confirming that antibodies produced in our cell-free system are comparable to those made in traditional cell-based systems in terms of binding efficiency, post-translational modifications, and functional activity, we can offer customers a powerful alternative during the discovery phase.”

In addition, the ability to swiftly optimize parameters—like heavy chain to light chain ratios or plasmid design—can accelerate the journey from bench to clinical trials, potentially transforming the landscape of antibody therapeutics.

LenioBio continues to drive forward the development of cell-free technologies, ensuring that cutting-edge research transforms into next-generation therapies. For more insights into the world of multispecific antibodies and the ALiCE® platform, explore our latest case study.