Home Analytical Chem Preparation and Characterization of Nanoliposomes for the Entrapment of Bioactive Hydrophilic Globular Proteins
Analytical Chem JoVE (Open Access) Citable · DOI

Preparation and Characterization of Nanoliposomes for the Entrapment of Bioactive Hydrophilic Globular Proteins

DOI: 10.3791/59900-v
What you'll learn
  • Prepare nanoliposomes using thin lipid film hydration method
  • Measure protein encapsulation efficiency and nanoparticle size
  • Characterize liposome stability and morphology by multiple techniques
  • Adapt protocol for hydrophobic or hydrophilic cargo encapsulation
Protocol

This study describes classical hydration using the thin lipid film method for nanoliposome preparation followed by nanoparticle characterization. A 47 kDa-hydrophilic and globular protein, tarin, is successfully encapsulated as a strategy to improve stability, avoid fast clearance, and promote controlled release. The method can be adapted to hydrophobic molecules encapsulation.

Difficulty
intermediate
Total time
~4-6 hours per batch (including drying, hydration, and initial characterization)

Steps

1
Prepare tarin liposomal nanocapsules via thin film hydration

Dissolve lipids in organic solvent, evaporate to form thin film, then hydrate with aqueous protein solution to generate nanoliposome suspension containing encapsulated tarin.

▶ 00:52
2
Determine protein encapsulation efficiency percentage

Quantify amount of tarin trapped within liposomes versus total protein using appropriate analytical method to assess encapsulation success.

▶ 04:24
3
Measure nanoliposome size and long-term stability

Use dynamic light scattering or similar technique to determine particle diameter and monitor size changes over storage time.

▶ 05:58
4
Characterize liposome morphology via electron microscopy

Visualize nanoliposome structure and confirm spherical morphology using transmission or scanning electron microscopy.

▶ 06:31
5
Interpret encapsulation and characterization results

Compile and analyze data from efficiency, size, stability, and morphology measurements to validate successful nanoliposome preparation.

▶ 08:29
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