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Innovative high-throughput SAXS methodologies based on photonic lab-on-a-chip sensors: application to macromolecular studies

Rodríguez-Ruiz, Isaac and Radajewski, Dimitri and Charton, Sophie and Pham, Van Nhat and Brennich, Martha and Pernot, Petra and Bonneté, Françoise and Teychené, Sébastien Innovative high-throughput SAXS methodologies based on photonic lab-on-a-chip sensors: application to macromolecular studies. (2017) Sensors, 17 (6). 1-12. ISSN 1424-8220

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Official URL: https://doi.org/10.3390/s17061266


The relevance of coupling droplet-based Photonic Lab-on-a-Chip (PhLoC) platforms and Small-Angle X-Ray Scattering (SAXS) technique is here highlighted for the performance of high throughput investigations, related to the study of protein macromolecular interactions. With this configuration, minute amounts of sample are required to obtain reliable statistical data. The PhLoC platforms presented in this work are designed to allow and control an effective mixing of precise amounts of proteins, crystallization reagents and buffer in nanoliter volumes, and the subsequent generation of nanodroplets by means of a two-phase flow. Spectrophotometric sensing permits a fine control on droplet generation frequency and stability as well as on concentration conditions, and finally the droplet flow is synchronized to perform synchrotron radiation SAXS measurements in individual droplets (each one acting as an isolated microreactor) to probe protein interactions. With this configuration, droplet physic-chemical conditions can be reproducibly and finely tuned, and monitored without cross-contamination, allowing for the screening of a substantial number of saturation conditions with a small amount of biological material. The setup was tested and validated using lysozyme as a model of study. By means of SAXS experiments, the proteins gyration radius and structure envelope were calculated as a function of protein concentration. The obtained values were found to be in good agreement with previously reported data, but with a dramatic reduction of sample volume requirements compared to studies reported in the literature.

Item Type:Article
HAL Id:hal-02134844
Audience (journal):International peer-reviewed journal
Uncontrolled Keywords:
Institution:French research institutions > Commissariat à l'Energie Atomique et aux énergies alternatives - CEA (FRANCE)
French research institutions > Centre National de la Recherche Scientifique - CNRS (FRANCE)
Université de Toulouse > Institut National Polytechnique de Toulouse - Toulouse INP (FRANCE)
Université de Toulouse > Université Toulouse III - Paul Sabatier - UT3 (FRANCE)
Other partners > European Molecular Biology Laboratory - EMBL (GERMANY)
Other partners > Université d'Avignon et des Pays de Vaucluse (FRANCE)
Laboratory name:
Deposited On:11 Oct 2018 09:37

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