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Measuring Enzymatic Stability by Isothermal Titration Calorimetry
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rhGALNS Enzyme Stability in Physiological Buffers: Implications for Sustained Release.

Samuel Ruesing1, Samuel Stealey1, Qi Gan2

  • 1Department of Biomedical Engineering, Saint Louis University, 3507 Lindell Blvd, St. Louis, MO, 63103, USA.

Applied Biochemistry and Biotechnology
|May 24, 2025
PubMed
Summary

Polyethylene glycol hydrogels can stabilize the enzyme N-acetylgalactosamine-6-sulfate sulfatase (GALNS) for treating Morquio A syndrome. Encapsulation within hydrogels improved enzyme stability compared to buffer solutions, enabling potential sustained release therapies.

Keywords:
Drug deliveryEnzyme replacement therapyLysosomal storage disorderMorquio APolyethylene glycol hydrogel

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Area of Science:

  • Biochemistry
  • Biomaterials Science
  • Genetic Disorders

Background:

  • Morquio A syndrome results from a deficiency in the GALNS enzyme, impairing glycosaminoglycan breakdown.
  • Current recombinant human GALNS (rhGALNS) treatments involve costly, time-consuming intravenous infusions with limited efficacy.
  • Developing a sustained-release injectable device could significantly improve patient quality of life.

Purpose of the Study:

  • To investigate the stability of rhGALNS in physiological buffers and within polyethylene glycol (PEG) hydrogels.
  • To assess the feasibility of using PEG hydrogels for sustained delivery of rhGALNS.

Main Methods:

  • rhGALNS stability was tested in phosphate-buffered saline (PBS) and acidic buffer.
  • Enzyme activity and hydrodynamic radius were measured after incubation in different conditions.
  • rhGALNS was encapsulated in PEG hydrogels, and its activity was monitored during release over 7 days.

Main Results:

  • rhGALNS activity decreased by 85% in PBS due to reversible inhibition, which was recoverable in acidic buffer.
  • Incubation in PBS for 3 days caused an irreversible 85% loss of activity, without altering hydrodynamic radius.
  • rhGALNS activity was retained upon encapsulation, with only a 20% decrease in specific activity over 7 days of release from the hydrogel.

Conclusions:

  • rhGALNS is unstable in physiological buffers like PBS, hindering sustained delivery.
  • Confinement within PEG hydrogels significantly enhances rhGALNS stability and activity retention.
  • PEG hydrogels show promise as a delivery system for sustained release of rhGALNS and other enzymes unstable in vivo.