Non-invasive detection of protein aggregation in biopharmaceuticals using TD-NMR and MRI

Abstract

With the rapid expansion of biotechnological drugs, ensuring their quality has become essential. Proper storage and transportation are critical to maintaining drug stability and efficacy. This study investigates protein aggregation, a major quality concern in biopharmaceuticals, using low-field NMR, Time Domain NMR and Magnetic Resonance Imaging (MRI), as a non-invasive, rapid alternative to conventional analytical methods. For the first time, MRI, T₂ relaxation times, and T₂ relaxation spectra were applied to detect aggregation in Humulin R, a low-concentration subcutaneous insulin. Aggregation was induced by heat and agitation to simulate improper handling. MRI effectively distinguished control and stressed samples, while T₂ relaxation measurements differentiated stress conditions. Additionally, T₂-Inverse Laplace Transform (ILT) indicated aggregate size, aligning with particle size analysis and validating the novel approach. These findings highlight the potential of low-field NMR for biopharmaceutical quality assessment.