Related Experiment Video
Updated: May 28, 2026
08:03
Study of Protein Dynamics via Neutron Spin Echo Spectroscopy
Published on: April 13, 2022
High-throughput small-angle X-ray scattering reveals effective structure factor transitions linked to
Lateefat Kalejaye1, I-En Wu1, Jia-Min Chu1
1Department of Chemical Engineering and Materials Science, Stevens Institute of Technology, Hoboken, New Jersey, USA.
Mabs
|May 27, 2026
Summary
A new high-throughput small-angle X-ray scattering (SAXS) method predicts high-concentration monoclonal antibody (mAb) viscosity using dilute solutions. This approach enables early risk assessment for antibody developability and manufacturing.
Area of Science:
- Biophysical Chemistry
- Protein Engineering
- Pharmaceutical Sciences
Background:
- High-concentration monoclonal antibody (mAb) formulations often face viscosity challenges due to protein-protein interactions, hindering manufacturing and delivery.
- Current viscosity measurements require large sample volumes and lack high-throughput capabilities, delaying early risk assessment during drug discovery.
Purpose of the Study:
- To develop and validate a high-throughput small-angle X-ray scattering (SAXS) protocol for early detection of mAb self-association at dilute concentrations.
- To enable predictive insights into high-concentration viscosity and assess antibody developability early in the discovery process.
Main Methods:
- Utilized synchrotron SAXS to analyze 21 mAbs across a range of concentrations (1-150 mg/mL) in histidine buffer (pH 6.0).
- Employed automated liquid handling and flow cells for high-throughput screening of 11 additional mAbs at 1-25 mg/mL.
- Analyzed low-q region transitions in the effective structure factor to identify interparticle interactions.
Main Results:
- Effective structure factor transitions indicative of interparticle interactions were observed below 25 mg/mL for a subset of mAbs.
- High-viscosity mAbs showed detectable low-q upturns at concentrations ≤10 mg/mL, while low-viscosity mAbs exhibited downturns.
- A classification criterion based on these transitions accurately predicted viscosity for all mAbs at 150 mg/mL.
Conclusions:
- The developed high-throughput SAXS protocol offers a scalable and sample-efficient alternative for early viscosity risk assessment of monoclonal antibody formulations.
- SAXS sensitivity to short-range attractions at lower concentrations than previously reported facilitates rational antibody engineering and improved developability.
- The comprehensive SAXS dataset provides a valuable resource for developing and validating models of intermolecular interactions in concentrated antibody solutions.
