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In Situ Measurements of Polypeptide Samples by Dynamic Light Scattering: Membrane Proteins, a Case Study

Part of the Methods in Molecular Biology book series (MIMB,volume 2208)


Integral membrane proteins are important drug targets that are critical in supporting many biological processes. Despite that, the study of their structure–function relationships remains a major goal in structural biology, yet progress has been hampered by inherent challenges in the production for stable and homogeneous protein samples. Dynamic light scattering provides a straightforward probe of protein quality in solution, particularly in relation to stability and aggregation. However, the necessity to use large amounts of sample and the low-throughput nature of the analysis remain as major bottlenecks of the technique.

Here, we present a protocol for dynamic light scattering measurements that are executed in a fully automated fashion for low-volume samples, in situ. The protocol offers a generic pre-screening method for downstream structural studies of biomolecules using higher-resolution approaches such as X-ray crystallography, electron microscopy, small-angle X-ray scattering, and NMR .

Key words

  • Dynamic light scattering
  • Membrane proteins and biophysics
  • Biomembrane interfaces
  • Biomolecules
  • Protein solubility and aggregation
  • Crystal nucleation
  • Monodispersity
  • Hydrodynamic radius

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We acknowledge funding from the United Kingdom’s Department of Business, Energy and Industrial Strategy (BEIS).

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Correspondence to Isabel Moraes .

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Kwan, T.O.C., Reis, R., Moraes, I. (2021). In Situ Measurements of Polypeptide Samples by Dynamic Light Scattering: Membrane Proteins, a Case Study. In: Ryadnov, M. (eds) Polypeptide Materials. Methods in Molecular Biology, vol 2208. Humana, New York, NY.

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0927-9

  • Online ISBN: 978-1-0716-0928-6

  • eBook Packages: Springer Protocols