Low-field NMR Works as a Rapid, Automatic, Non-Invasive and Wide-Scale Coverage Technique for Aggregates Indication...

Protein aggregation is challenging for biopharmaceutical drug, because it affects the stability of protein formulations in real-time. However, current techniques for protein aggregate indication meet a number of limitations including limited aggregate size range, complex pre-treatments and lack of chromatographic approaches. Herein, a rapid, automatic, non-invasive and wide-scale coverage technique for aggregates indication is developed to overcome these challenges. Firstly, the response of low-field nuclear magnetic resonance (LF-NMR) to the aggregates is explored by making a comparison with certain established techniques. LF-NMR achieves a high sensitivity of water proton transverse relaxation rate (R₂ of H₂O, hereinafter referred as R₂(H₂O)) to protein aggregates from nanometer to micrometer. Then, the quantitative relationship between R₂(H₂O) and aggregates is investigated furtherly. R₂(H₂O) could serve as an all-size coverage protein aggregates indicator during development. As a non-invasive method, LF-NMR does not need any sample handling. It takes only 44 s for one test, and saves a lot of manpower, materials and costs. Compared with other established analytical techniques, the technique developed here could be a powerful tool for a rapid, automatic, non-invasive and wide-scale coverage technique for aggregates indication in biomacromolecule development.