Adeno-Associated Virus-like Particles’ Response to pH Changes as Revealed by nES-DMA
Gas-phase electrophoresis on a nano-Electrospray Gas-phase Electrophoretic Mobility Molecular Analyzer (nES GEMMA) separates single-charged, native analytes according to the surface-dry particle size. A volatile electrolyte, often ammonium acetate, is a prerequisite for electrospraying. Over the yea...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-06-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/15/6/1361 |
| _version_ | 1827735356003319808 |
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| author | Samuele Zoratto Thomas Heuser Gernot Friedbacher Robert Pletzenauer Michael Graninger Martina Marchetti-Deschmann Victor U. Weiss |
| author_facet | Samuele Zoratto Thomas Heuser Gernot Friedbacher Robert Pletzenauer Michael Graninger Martina Marchetti-Deschmann Victor U. Weiss |
| author_sort | Samuele Zoratto |
| collection | DOAJ |
| description | Gas-phase electrophoresis on a nano-Electrospray Gas-phase Electrophoretic Mobility Molecular Analyzer (nES GEMMA) separates single-charged, native analytes according to the surface-dry particle size. A volatile electrolyte, often ammonium acetate, is a prerequisite for electrospraying. Over the years, nES GEMMA has demonstrated its unique capability to investigate (bio-)nanoparticle containing samples in respect to composition, analyte size, size distribution, and particle numbers. Virus-like particles (VLPs), being non-infectious vectors, are often employed for gene therapy applications. Focusing on adeno-associated virus 8 (AAV8) based VLPs, we investigated the response of these bionanoparticles to pH changes via nES GEMMA as ammonium acetate is known to exhibit these changes upon electrospraying. Indeed, slight yet significant differences in VLP diameters in relation to pH changes are found between empty and DNA-cargo-filled assemblies. Additionally, filled VLPs exhibit aggregation in dependence on the applied electrolyte’s pH, as corroborated by atomic force microscopy. In contrast, cryogenic transmission electron microscopy did not relate to changes in the overall particle size but in the substantial particle’s shape based on cargo conditions. Overall, we conclude that for VLP characterization, the pH of the applied electrolyte solution has to be closely monitored, as variations in pH might account for drastic changes in particles and VLP behavior. Likewise, extrapolation of VLP behavior from empty to filled particles has to be carried out with caution. |
| first_indexed | 2024-03-11T01:49:53Z |
| format | Article |
| id | doaj.art-3d32ed71de5e4fa9bae03f0dd29d73b1 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-11T01:49:53Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-3d32ed71de5e4fa9bae03f0dd29d73b12023-11-18T13:02:45ZengMDPI AGViruses1999-49152023-06-01156136110.3390/v15061361Adeno-Associated Virus-like Particles’ Response to pH Changes as Revealed by nES-DMASamuele Zoratto0Thomas Heuser1Gernot Friedbacher2Robert Pletzenauer3Michael Graninger4Martina Marchetti-Deschmann5Victor U. Weiss6Institute of Chemical Technologies and Analytics, TU Wien, A-1060 Vienna, AustriaElectron Microscopy Facility, Vienna BioCenter Core Facilities GmbH, A-1030 Vienna, AustriaInstitute of Chemical Technologies and Analytics, TU Wien, A-1060 Vienna, AustriaPharmaceutical Sciences, Baxalta Innovations GmbH (Part of Takeda), A-1221 Vienna, AustriaPharmaceutical Sciences, Baxalta Innovations GmbH (Part of Takeda), A-1221 Vienna, AustriaInstitute of Chemical Technologies and Analytics, TU Wien, A-1060 Vienna, AustriaInstitute of Chemical Technologies and Analytics, TU Wien, A-1060 Vienna, AustriaGas-phase electrophoresis on a nano-Electrospray Gas-phase Electrophoretic Mobility Molecular Analyzer (nES GEMMA) separates single-charged, native analytes according to the surface-dry particle size. A volatile electrolyte, often ammonium acetate, is a prerequisite for electrospraying. Over the years, nES GEMMA has demonstrated its unique capability to investigate (bio-)nanoparticle containing samples in respect to composition, analyte size, size distribution, and particle numbers. Virus-like particles (VLPs), being non-infectious vectors, are often employed for gene therapy applications. Focusing on adeno-associated virus 8 (AAV8) based VLPs, we investigated the response of these bionanoparticles to pH changes via nES GEMMA as ammonium acetate is known to exhibit these changes upon electrospraying. Indeed, slight yet significant differences in VLP diameters in relation to pH changes are found between empty and DNA-cargo-filled assemblies. Additionally, filled VLPs exhibit aggregation in dependence on the applied electrolyte’s pH, as corroborated by atomic force microscopy. In contrast, cryogenic transmission electron microscopy did not relate to changes in the overall particle size but in the substantial particle’s shape based on cargo conditions. Overall, we conclude that for VLP characterization, the pH of the applied electrolyte solution has to be closely monitored, as variations in pH might account for drastic changes in particles and VLP behavior. Likewise, extrapolation of VLP behavior from empty to filled particles has to be carried out with caution.https://www.mdpi.com/1999-4915/15/6/1361AAV8VLPnES GEMMADMAcryo-TEMgene therapy |
| spellingShingle | Samuele Zoratto Thomas Heuser Gernot Friedbacher Robert Pletzenauer Michael Graninger Martina Marchetti-Deschmann Victor U. Weiss Adeno-Associated Virus-like Particles’ Response to pH Changes as Revealed by nES-DMA Viruses AAV8 VLP nES GEMMA DMA cryo-TEM gene therapy |
| title | Adeno-Associated Virus-like Particles’ Response to pH Changes as Revealed by nES-DMA |
| title_full | Adeno-Associated Virus-like Particles’ Response to pH Changes as Revealed by nES-DMA |
| title_fullStr | Adeno-Associated Virus-like Particles’ Response to pH Changes as Revealed by nES-DMA |
| title_full_unstemmed | Adeno-Associated Virus-like Particles’ Response to pH Changes as Revealed by nES-DMA |
| title_short | Adeno-Associated Virus-like Particles’ Response to pH Changes as Revealed by nES-DMA |
| title_sort | adeno associated virus like particles response to ph changes as revealed by nes dma |
| topic | AAV8 VLP nES GEMMA DMA cryo-TEM gene therapy |
| url | https://www.mdpi.com/1999-4915/15/6/1361 |
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