Probing in situ higher order structures of monoclonal antibodies at water-air and water-oil interfaces via high-field nuclear magnetic resonance spectroscopy for viral infections

通过高场核磁共振波谱技术在水-空气和水-油界面原位探测单克隆抗体的高阶结构以检测病毒感染

基本信息

批准号：
10593377
负责人：
Hadi Mohammadigoushki
金额：
$ 22.49万
依托单位：
FLORIDA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-17 至 2025-07-31
项目状态：
未结题

项目摘要

PROJECT SUMMARY/ABSTRACT Monoclonal antibodies (mAbs) represent an important class of biologic therapeutics that can treat COVID-19, cancer and other infectious diseases. Despite their promising potential, pro- cessing, storage and/or administration of mAbs into patients is challenging because the presence of hydrophobic interfaces during processing and administration (air entrapment in the IV bags or the oil-water interface at the interior of syringes) may promote mAb adsorption to such hy- drophobic interfaces. If mAbs change their native (folded) higher order structures (HOS) upon adsorption to these interfaces, their quality, safety and efficacy will be affected, posing immuno- genicity risks to already susceptible patients. The first step in mitigating these risks is to evaluate the in situ HOS of mAbs (whether folded or unfolded) at hydrophobic interfaces. Determining the in situ structure of mAbs at such interfaces has been a major challenge due to limitations of bulk scale or scattering-based microstructural probing techniques. In this program, we will go beyond such limits and use a combination of a unique molecular probing technique based on NMR spec- troscopy and dynamic surface tensiometry to resolve the details of mAbs HOS and adsorption kinetics at hydrophobic interfaces. In particular, by using high-field spatially and spectrally re- solved NMR spectroscopy that is uniquely available to use through National High Magnetic Field Laboratory, we will assess dynamically 1) the in situ HOS of pure mAbs at hydrophobic interfaces, and 2) nature of their associations with surfactants at interfaces. We will perform tensiometry along with NMR spectroscopy on pure mAbs, isotopically labeled mAbs and mAbs/surfactant combinations at hydrophobic interfaces. We will measure a) dynamic surface tension, b) spa- tially localized chemical shifts in 1D 1H and 2D 1H-13C NMR spectra, c) diffusion coefficients of the mAbs, and d) T2 relaxation of mAbs in the bulk and at the interface under different conditions (e.g., various mAbs and surfactant concentrations, solution pH and ionic strengths). By compar- ing the results of the bulk and interface in terms of metrics (a-d), the team will determine if the native HOS of mAbs has been altered by adsorption to hydrophobic interfaces or their associa- tions with surfactants. The outcome of this study will provide the first mechanistic understanding of mAbs HOS at hydrophobic interfaces. Additionally, the knowledge gained from this research is essential in developing a framework to mitigate mAbs adsorption to hydrophobic interfaces, which can be subsequently utilized to improve efficacious mAb deployment for patients. 1

项目摘要/摘要单克隆抗体（mAb）代表一类重要的生物疗法，可以治疗Covid-19，癌症和其他传染病。尽管他们有希望的潜力，但由于存在加工和给药过程中疏水界面的或注射器内部的油水界面）可能会促进对此类Hy-的mAb吸附疏忽界面。如果mabs改变其本地（折叠）高阶结构（HOS）吸附到这些接口，它们的质量，安全性和功效将受到影响，从而带来免疫已经易感患者的统一风险。减轻这些风险的第一步是评估在疏水界面处mab的原位hos（无论是折叠还是展开）。确定由于批量的局限基于比例或散射的微结构探测技术。在这个程序中，我们将超越这种限制并结合了基于NMR Spec-的独特分子探测技术 Troscopy和动态表面张力仪，以解决mAbs HOS的细节和吸附的细节疏水界面的动力学。特别是，通过在空间和光谱上使用高场已解决的NMR光谱法可用于通过国家高磁场使用实验室，我们将动态评估1）在疏水界面处纯MAB的原位HOS， 2）它们与界面上表面活性剂的关联性质。我们将执行张力计以及纯MAB上的NMR光谱，同位素标记的mAb和mAb/表面活性剂疏水界面的组合。我们将测量a）动态表面张力，b）spa- 在1d 1h和2d 1h-13c NMR光谱中的局部定位化学位移，c）扩散系数的扩散系数 mabs，在不同条件下的散装和界面中mAb的d）t2松弛（例如，各种mAb和表面活性剂浓度，溶液pH和离子强度）。通过比较根据指标（A-D）的批量和界面的结果，团队将确定是否是否通过吸附到疏水界面或它们的相关性，MAB的天然HOS已改变了表面活性剂。这项研究的结果将提供第一个机械理解在疏水界面处的mabs hos。此外，这项研究所获得的知识对于开发一个框架以减轻mAb的吸附到疏水界面，至关重要，随后可以用来改善患者的有效MAB部署。 1