No ordinary proteins: Adsorption and molecular orientation of monoclonal antibodies

The interaction of monoclonal antibodies (mAbs) with air/water interfaces plays a crucial role in their overall stability in solution. We aim to understand this behavior using pendant bubble measurements to track the dynamic tension reduction and x-ray reflectivity to obtain the electron density profiles (EDPs) at the surface. Native immunoglobulin G mAb is a rigid molecule with a flat, Y shape, and simulated EDPs are obtained by rotating a homology construct at the surface. Comparing simulations with experimental EDPs, we obtain surface orientation probability maps showing mAbs transition from flat-on Y-shape configurations to side-on or end-on configurations with increasing concentration. The modeling also shows the presence of. sheets at the surface. Overall, the experiments and the homology modeling elucidate the orientational phase space during different stages of adsorption of mAbs at the air/water interface. These finding will help define new strategies for the manufacture and storage of antibody-based therapeutics.

Automated summary

The interaction of monoclonal antibodies with air/water interfaces is crucial for their stability in solution, which can be studied using pendant bubble measurements and x-ray reflectivity to obtain electron density profiles at the surface. By comparing simulations and experimental data, it is shown that mAbs transition from flat-on Y-shape configurations to side-on or end-on configurations at the interface, with the presence of β sheets. This understanding can lead to new strategies for the manufacture and storage of antibody-based therapeutics.