Nano Differential Scanning Fluorimetry as a Rapid Stability Assessment Tool in the Nanoformulation of Proteins.

Content Provider	Europe PMC
Author	Lisina, Sofia Inam, Wali Huhtala, Mikko Howaili, Fadak Zhang, Hongbo Rosenholm, Jessica M.
Editor	Grohganz, Holger
Copyright Year	2023
Abstract	The development and production of innovative protein-based therapeutics is a complex and challenging avenue. External conditions such as buffers, solvents, pH, salts, polymers, surfactants, and nanoparticles may affect the stability and integrity of proteins during formulation. In this study, poly (ethylene imine) (PEI) functionalized mesoporous silica nanoparticles (MSNs) were used as a carrier for the model protein bovine serum albumin (BSA). To protect the protein inside MSNs after loading, polymeric encapsulation with poly (sodium 4-styrenesulfonate) (NaPSS) was used to seal the pores. Nano differential scanning fluorimetry (NanoDSF) was used to assess protein thermal stability during the formulation process. The MSN-PEI carrier matrix or conditions used did not destabilize the protein during loading, but the coating polymer NaPSS was incompatible with the NanoDSF technique due to autofluorescence. Thus, another pH-responsive polymer, spermine-modified acetylated dextran (SpAcDEX), was applied as a second coating after NaPSS. It possessed low autofluorescence and was successfully evaluated with the NanoDSF method. Circular dichroism (CD) spectroscopy was used to determine protein integrity in the case of interfering polymers such as NaPSS. Despite this limitation, NanoDSF was found to be a feasible and rapid tool to monitor protein stability during all steps needed to create a viable nanocarrier system for protein delivery.
Journal	Pharmaceutics
Volume Number	15
PubMed Central reference number	PMC10223687
Issue Number	5
PubMed reference number	37242715
e-ISSN	19994923
DOI	10.3390/pharmaceutics15051473
Language	English
Publisher	MDPI
Publisher Date	2023-05-11
Access Restriction	Open
Rights License	Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). © 2023 by the authors.
Subject Keyword	protein stability bovine serum albumin microfluidics protein encapsulation mesoporous silica nanoparticles nano differential scanning fluorimetry
Content Type	Text
Resource Type	Article
Subject	Pharmaceutical Science