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Lifting the Concentration Limit of Mass Photometry by PEG Nanopatterning.


ABSTRACT: Mass photometry (MP) is a rapidly growing optical technique for label-free mass measurement of single biomolecules in solution. The underlying measurement principle provides numerous advantages over ensemble-based methods but has been limited to low analyte concentrations due to the need to uniquely and accurately quantify the binding of individual molecules to the measurement surface, which results in diffraction-limited spots. Here, we combine nanoparticle lithography with surface PEGylation to substantially lower surface binding, resulting in a 2 orders of magnitude improvement in the upper concentration limit associated with mass photometry. We demonstrate the facile tunability of degree of passivation, enabling measurements at increased analyte concentrations. These advances provide access to protein-protein interactions in the high nanomolar to low micromolar range, substantially expanding the application space of mass photometry.

SUBMITTER: Kratochvil J 

PROVIDER: S-EPMC11342371 | biostudies-literature | 2024 Jul

REPOSITORIES: biostudies-literature

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Lifting the Concentration Limit of Mass Photometry by PEG Nanopatterning.

Kratochvíl Jiří J   Asor Roi R   Helmi Seham S   Struwe Weston B WB   Kukura Philipp P  

Nano letters 20240701 33


Mass photometry (MP) is a rapidly growing optical technique for label-free mass measurement of single biomolecules in solution. The underlying measurement principle provides numerous advantages over ensemble-based methods but has been limited to low analyte concentrations due to the need to uniquely and accurately quantify the binding of individual molecules to the measurement surface, which results in diffraction-limited spots. Here, we combine nanoparticle lithography with surface PEGylation t  ...[more]

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