# Technology Development for High-Throughput Biomolecular CryoSAXS

> **NIH NIH R01** · CORNELL UNIVERSITY · 2021 · $363,473

## Abstract

Project Summary/Abstract
 In the last fifteen years, small-angle X-ray scattering (SAXS) has become a key tool in academic,
government, and commercial research and development for probing the structure and function of proteins,
nucleic acids, and macromolecular complexes. SAXS yields precise but low resolution structural information
from biomolecules and complexes in solution, without the need to crystallize or label the biomolecule, regardless
of molecular weight or the extent of molecular order. Sophisticated and easy-to-use data analysis suites enable
rapid interpretation of SAXS profiles, yielding information ranging from molecular radius of gyration to structural
envelopes and ensembles. Most synchrotron sources have dedicated bioSAXS beam lines that allow
measurement of hundreds of samples per day, but ongoing efforts to improve their throughput have not kept
pace with the rapid expansion in user demand.
 Currently, all bioSAXS is performed on samples at or near room temperature. Due to biomolecular
aggregation and degradation between initial expression/purification and SAXS measurements and to radiation
damage by the illuminating X-rays, large volumes of sample are required per measurement. Sample cells must
be loaded and then thoroughly cleaned between each measurement, so data collection duty cycles are very low.
Large sample volume requirements, low measurement duty cycles, and high user demand are critical bottlenecks
in the continued expansion of bioSAXS, especially for high-throughput parameter and ligand interaction
screening and for study of difficult to produce proteins or complexes, applications in which bioSAXS may have
the greatest impact on human health.
 Building upon our recent demonstration experiments, this project aims to develop technology and methods
for high-throughput SAXS on biomolecular samples at cryogenic temperatures. As with cryocrystallography,
cryoSAXS should require much smaller sample volumes per measurement, allow sample preparation in the
home lab immediately after purification, easy sample storage and shipping, and automated high-throughput data
collection with duty cycles approaching 50%. This will enable dramatically more efficient use of both
biomolecules and synchrotron beam time, and significantly expand the potential scope of bioSAXS studies. Key
aspects of this technology to be developed are (1) fixed and reproducible path length sample cells and sample
cell arrays that enable rapid, homogeneous sample cooling; (2) tools for sample cooling, storage, shipment, and
high-throughput handling based on those for high-throughput cryocrystallography; (3) screens of contrast-
maximizing cryoprotective buffers of known cryogenic electron density that minimize thermomechanical stresses;
(4) measurement-based modeling of hydration layers at cryogenic temperatures, needed for interpretation of
cryoSAXS profiles; and (5) extensions of this technology to SAXS studies on biomolecules in supercooled liquid
buffers ...

## Key facts

- **NIH application ID:** 10115760
- **Project number:** 5R01GM127528-04
- **Recipient organization:** CORNELL UNIVERSITY
- **Principal Investigator:** Robert E. Thorne
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $363,473
- **Award type:** 5
- **Project period:** 2018-03-01 → 2023-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10115760

## Citation

> US National Institutes of Health, RePORTER application 10115760, Technology Development for High-Throughput Biomolecular CryoSAXS (5R01GM127528-04). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10115760. Licensed CC0.

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