Abstract：
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.
摘要：
蛋白质聚集是生物制药药物的挑战，因为它会影响实时蛋白质制剂的稳定性。然而,目前用于蛋白质聚集体适应症的技术满足许多限制，包括有限的聚集体大小范围，复杂的预处理和缺乏色谱方法。在这里，一个快速的，自动,为了克服这些挑战,开发了非侵入性和广泛覆盖聚合适应症的技术.首先,通过与某些已建立的技术进行比较，探索了低场核磁共振（LF-NMR）对聚集体的响应。LF-NMR实现了水质子横向弛豫率的高灵敏度（H2O的R2，以下称为R2（H2O）），从纳米到微米的蛋白质聚集体。然后，进一步研究了R2（H2O）与聚集体之间的定量关系。R2（H2O）可以在发育过程中充当全尺寸覆盖蛋白聚集体指标。作为一种非侵入性方法，LF-NMR不需要任何样品处理。一次测试只需要44秒，节省了大量的人力，材料和成本。与其他既定的分析技术相比，这里开发的技术可能是一个强大的工具，自动,生物大分子发育中聚集体适应症的非侵入性和广泛覆盖技术。
