Reengineering thermal field-flow fractionation for biocompatibility: A novel approach for characterizing binding affinity and microstructural dynamics between biopharmaceutical nanostructures and active pharmaceutical ingredients

• Applicant: Dr. Upenyu Muza
• Subject Area: Analytical Chemistry ; Biochemistry ; Biophysics
• Term: since 2024
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 551623767

BIO-TF3 aims to excel beyond the state-of-the-art in the characterization of biopharmaceutical nanostructures (BPNs) and their interactions with active pharmaceutical ingredients (APIs) by integrating two cutting-edge techniques: microscale thermophoresis (MST) and thermal field-flow fractionation (ThFFF). Currently, MST provides precise binding affinity measurements but lacks separative capabilities and statistical assessments of property distributions. ThFFF, on the other hand, is capable of separating complex nanostructures, but has not been applied to BPNs. By combining these techniques into a singular analytical procedure (ThFFF – MST), BIO-TF3 seeks to overcome the inherent limitations and provide comprehensive insights into both binding affinity and microstructural dynamics of BPN - API complexes. By accomplishing the following objectives, BIO-TF3 aims to advance the understanding of BPN - API interactions, paving the way for more effective targeted drug delivery and advanced therapeutics: (i). Develop a biocompatible ThFFF system capable of separative MST to analyze sensitive biological entities like BPNs. This involves electroplating the ThFFF channel with gold for chemical stability and hyphenating it with fluorescence detection to enable separative MST; (ii) Implement an innovative data analysis framework called 3-dimensional correlation ThFFF (3DCoThFFF) to enhance the interpretation of complex datasets generated from ThFFF – MST; (iii) Characterize various BPNs (such as extracellular vesicles, solid lipid nanoparticles, and functionalized polymersomes) to demonstrate the efficacy of ThFFF – MST, especially in cancer therapeutics; (iv) Integrate ThFFF with small-angle X-ray scattering (SAXS) to gain high-resolution insights into the microstructural dynamics of BPNs under physiological conditions; (v) Utilize multiple detectors downstream of ThFFF – MST to provide complementary analytics and validation datasets within a single measurement, ensuring accuracy and reliability; (vi) Address the limitations of conventional techniques like batch-wise MST by providing separative capabilities and self-validation within ThFFF – MST.

• DFG Programme: Research Grants
• International Connection: Bulgaria, Portugal, Sweden
• Cooperation Partners: Professor Lars Nilsson, Ph.D.; Professor António José Braga Osório Gomes Salgado, Ph.D.; Professor Christo Tzachev, Ph.D.