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Size-Exclusion Chromatography01:08

Size-Exclusion Chromatography

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In size-exclusion chromatography (SEC), also known as molecular-exclusion or gel-permeation chromatography, molecules are separated based on their sizes. This technique is important for separating large molecules such as polymers and biomolecules. The two classes of micron-sized stationary phases encountered in SEC are silica particles and cross-linked polymer resin beads. Both materials are porous, but their pore sizes vary significantly.
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Cell separation was first achieved in 1964 by S. H. Seal, who separated large tumor cells from the smaller blood cells using filtration. Two years later, Pohl and Hawk performed experiments on how cells respond differently to a nonuniform electric field based on the cell type. Such observations were the inception of cell separation methods, which allow isolating a single cell type from a heterogeneous sample.
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Optimizing chromatographic separations is crucial for obtaining clean separations in a minimum amount of time. Optimization is required for several factors, including kinetic effects related to band broadening, plate height, capacity factor, and separation factor.
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Centrifugation is a separation technique based on differences in density or size. It is commonly used to separate solids from aqueous interferents. During centrifugation, the sample is placed in centrifugation tubes and spun at high angular velocity, which allows centrifugal force to act differentially on the different densities or masses of the components. After spinning, the supernatant liquid is decanted. Depending on the specific application, either the pellet or the supernatant is retained...
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Spatial Separation of Molecular Conformers and Clusters
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Phase separation and critical size in molecular sorting.

Elisa Floris1, Andrea Piras2, Francesco Saverio Pezzicoli3

  • 1Institute of Condensed Matter Physics and Complex Systems, Department of Applied Science and Technology, Politecnico di Torino, Corso Duca degli Abruzzi 24, 10129 Torino, Italy.

Physical Review. E
|November 18, 2022
PubMed
Summary
This summary is machine-generated.

Molecular sorting in eukaryotic cells relies on phase separation and membrane bending. Efficiency is maximized when the number of sorting domains is minimized, with a critical size defining domain fate.

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Area of Science:

  • Cell biology
  • Biophysics
  • Physical chemistry

Background:

  • Molecular sorting concentrates chemical factors in cell membrane subregions.
  • A recent theory links sorting to phase separation and membrane bending, forming lipid vesicles.
  • The critical size of phase-separated domains is key to sorting efficiency.

Purpose of the Study:

  • To explore the implications of domain critical size in molecular sorting theory.
  • To investigate the link between domain fate (productive vs. unproductive) and phase separation theory.
  • To validate theoretical predictions with numerical simulations and experimental data.

Main Methods:

  • Phenomenological theory of molecular sorting.
  • Numerical simulations of a lattice-gas model.
  • Analysis of experimental data on sorting domains.

Main Results:

  • Sorting efficiency is highest when the number of sorting domains is minimized.
  • An operational definition for the critical size of sorting domains is proposed.
  • Simulations align with experimental data on productive and unproductive domains.

Conclusions:

  • Molecular sorting is likely driven by phase separation processes.
  • The critical size of domains is a crucial parameter for efficient molecular distillation.
  • The proposed theory and simulation model provide a framework for analyzing experimental sorting data.