Challenges in estimating the encapsulation efficiency of proteins in polymersomes - Which is the best method?

Polymersomes are nanometric vesicles that can encapsulate large and hydrophilic biomolecules, such as proteins, in the aqueous core. Data in literature show large variation in encapsulation efficiency (%EE) values depending on the method used for calculation. We investigated different approaches (di...

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Bibliographic Details
Published inBrazilian Journal of Pharmaceutical Sciences Vol. 59
Main Authors Muso-Cachumba, Jorge Javier, Ruiz-Lara, Grace, Monteiro, Gisele, Rangel-Yagui, Carlota de Oliveira
Format Journal Article
LanguageEnglish
Published Sao Paulo Universidade de Sao Paulo Faculdade de Ciencias 01.01.2023
Universidade de São Paulo, Faculdade de Ciências Farmacêuticas
Universidade de São Paulo
Subjects
Biomolecules encapsulation
Drug delivery systems
Efficiency
Encapsulation efficiency
Encyclopedias
Enzymes
Internet
Nanomaterials
Nanoparticles
PHARMACOLOGY & PHARMACY
Poloxamer
Polymersomes
Proteins
Spectrum analysis
Vesicle disruption
United States > US
Encapsulation efficiency
Biomolecules encapsulation
Polymersomes
Vesicle disruption
Poloxamer
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Summary:Polymersomes are nanometric vesicles that can encapsulate large and hydrophilic biomolecules, such as proteins, in the aqueous core. Data in literature show large variation in encapsulation efficiency (%EE) values depending on the method used for calculation. We investigated different approaches (direct and indirect) to quantify the %EE of different proteins (catalase, bovine serum albumin-BSA, L-asparaginase and lysozyme) in Pluronic L-121 polymersomes. Direct methods allow quantification of the actual payload of the polymersomes and indirect methods are based on the quantification of the remaining non-encapsulated protein. The protein-loaded polymersomes produced presented approximately 152 nm of diameter (PDI ~ 0.4). Higher %EE values were obtained with the indirect method (up to 25%), attributed to partial entanglement of free protein in the polymersomes poly(Ethylene Glycol) corona. For the direct methods, vesicles were disrupted with chloroform or proteins precipitated with solvents. Reasonable agreement was found between the two protocols, with values up to 8%, 6%, 17.6% and 0.9% for catalase, BSA, L-asparaginase and lysozyme, respectively. We believe direct determination is the best alternative to quantify the %EE and the combination of both protocols would make results more reliable. Finally, no clear correlation was observed between protein size and encapsulation efficiency.
ISSN:2175-9790
1984-8250
2175-9790
DOI:10.1590/s2175-97902023e23365