Matthew Eddy

Increases the Stability and Robustness of the Galectin-3C Protein

PEGylation of the carbohydrate recognition domain of human Galectin-3 (Gal3C) increases the stability and robustness of the protein, overcoming limitations hindering its biologic therapeutic use. Biologics are highly effective, targeted treatments for varying diseases, constituting approximately 30% of drugs approved by the FDA in the past five years. Galectin-3 (Gal3C) has well-documented roles in multiple disease areas, including cancer, endometriosis and arthritis. It is a potential biologic useful in cancer treatment, inhibiting tumor growth in breast cancer animal models and enhancing the activity of anti-cancer compounds. Additional applications have been explored for the treatment of endometriosis and arthritis. However, Gal3C and other biologics are rapidly cleared from the body. Despite profound impacts on numerous medical fields, the need for proteins to survive and have a specific function in the challenging environment inside the human body limits the practical application of biologics. Gal3C suffers from clearance and competition with endogenous Gal3C following administration, limiting its utility. A promising approach for overcoming these challenges is PEGylation, polymer-based protein engineering.

PEGylation is a widely used approach in industrial and pharmaceutical applications. Since approving the first PEGylated protein drug in 1990, nearly 30 FDA-approved, PEGylated drugs are in the clinic treating an array of illnesses, including numerous successful anti-cancer therapeutics. PEGylation of a protein should preserve or enhance the activity of a protein drug while providing improved robustness. However, clear criteria do not exist for predicting the effects of PEGylation on protein properties, hindering its broader application.

Researchers at the University of Florida have developed a PEGylated form of Gal3C, increasing its stability and robustness. The PEGylation of Gal3C increases its thermal stability while maintaining proper formation and biological activity at physiological temperatures, allowing its potential as a cancer biologic to be tested and demonstrating methods for producing such proteins with high purity and at relatively large laboratory scales.

Application

PEGylated Galectin-3 (Gal3C) protein increases protein stability and robustness in the human body, improving its thermal stability and maintaining its proper formation and biological activity

Advantages

• Improves the thermal stability of Galectin-3 (Gal-3C), increasing the ability to study Gal3C’s use as a potential biologic for cancer treatment
• Provides a path for rationally designing protein-polymer conjugates, enabling the prediction of properties for these for the conjugates

Technology

The PEGylation of the Galectin-3 protein improves its biologic use, modulating its biophysical properties and improving stability while maintaining its biological activity. mPEG-maleimide (PEG) is added to Galectin-3 (Gal3C) protein via thiol-Michael conjugation to an extrinsic cysteine. The Gal3C-PEG conjugate forms a stable thermal unfolding intermediate when heated, increasing its thermal stability. This insight provides a method of designing protein-polymer conjugates suitable for therapeutics with high specificity, formulation and storage stability, and biocompatibility and utilizes Galectin-3C as a cancer biologic.