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SDS-PAGE

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Gel electrophoresis is a method that separates biological macromolecules like nucleic acids or proteins by forcing them to pass through a gel matrix under an electric field.
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Combining Non-reducing SDS-PAGE Analysis and Chemical Crosslinking to Detect Multimeric Complexes Stabilized by Disulfide Linkages in Mammalian Cells in Culture

Combining Non-reducing SDS-PAGE Analysis and Chemical Crosslinking to Detect Multimeric Complexes Stabilized by Disulfide Linkages in Mammalian Cells in Culture

Published on: May 2, 2019

Disulfide-Cross-Linked Tetra-PEG Gels.

Zhao Meng^1,2, Lucas Löser³, Kay Saalwächter³

¹Institut des Matériaux and Institut des Sciences et Ingénierie Chimiques, Laboratoire des Polymères, École Polytechnique Fédérale de Lausanne (EPFL), Bâtiment MXD, Station 12, CH-1015 Lausanne, Switzerland.

|April 15, 2024

Summary

This study explores creating disulfide-cross-linked polymer gels using self-reactive poly(ethylene glycol) macromers. Network structure depends on macromer concentration, offering potential for reduction-sensitive hydrogels in biomedical applications.

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Area of Science:

Polymer Chemistry
Materials Science
Biomedical Engineering

Background:

Polymer gels with defined topologies are often made using specific cross-linking methods.
Four-arm star-shaped poly(ethylene glycol) (Tetra-PEG) is a versatile precursor for network synthesis.
Traditional Tetra-PEG gels use heterocomplementary reactive precursors.

Purpose of the Study:

To investigate the formation and structure of disulfide-cross-linked Tetra-PEG gels using self-reactive thiol-end functional Tetra-PEG macromers.
To understand how macromer concentration influences network connectivity and topology.
To evaluate the potential of these disulfide-cross-linked gels as reduction-sensitive hydrogels.

Main Methods:

Synthesis of disulfide-cross-linked Tetra-PEG gels from thiol-end functional Tetra-PEG macromers.
Characterization of gel structure using multiple-quantum NMR (MQ-NMR) spectroscopy.
Analysis of network connectivity and topology via small-angle neutron scattering (SANS) experiments.

Main Results:

Network connectivity strongly depends on the concentration of the thiol-end functional Tetra-PEG macromer.
Gels synthesized below the overlap concentration exhibit a higher proportion of defect connectivity motifs (loops, dangling ends).
Above the overlap concentration, disulfide-cross-linked gels show similar single-link connectivities to amide-cross-linked Tetra-PEG gels.

Conclusions:

Self-reactive thiol-end functional Tetra-PEG macromers can form disulfide-cross-linked gels with tunable network structures.
Macromer concentration is a critical parameter controlling gel topology and defect formation.
The susceptibility of disulfide bonds to reductive cleavage makes these gels promising for biomedical applications requiring sensitivity to reduction.