# 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

> **NIH NIH R21** · FLORIDA STATE UNIVERSITY · 2023 · $224,875

## Abstract

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

## Key facts

- **NIH application ID:** 10593377
- **Project number:** 1R21AI163988-01A1
- **Recipient organization:** FLORIDA STATE UNIVERSITY
- **Principal Investigator:** Hadi Mohammadigoushki
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $224,875
- **Award type:** 1
- **Project period:** 2023-08-17 → 2025-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10593377

## Citation

> US National Institutes of Health, RePORTER application 10593377, 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 (1R21AI163988-01A1). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10593377. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*