The proportion of full and empty capsids represents a critical quality attribute of adeno-associated virus (AAV)-based therapeutics. In this study, pH-gradient anion exchange chromatography was utilized for the separation of full and empty capsid species. The developed method allowed for applicability to multiple AAV serotypes and facilitated subsequent mass spectrometric detection of intact AAVs. This is the first study demonstrating generic applicability as well as mass spectrometric compatibility, allowing for a more sophisticated analysis of AAV-based gene therapy and paving the way for future developments in the field.
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http://dx.doi.org/10.1039/d3ay01560b | DOI Listing |
J Virol
December 2024
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts, USA.
Unlabelled: Flavivirus envelope (E) and precursor M (prM) proteins, when ectopically expressed, assemble into empty, virus-like particles (VLPs). Cleavage of prM to M and loss of the pr fragment converts the VLPs from immature to mature particles, mimicking a similar maturation of authentic virions. Most of the VLPs obtained by prM-E expression are smaller than virions; early, low-resolution cryo-EM studies suggested a simple, 60-subunit, icosahedral organization.
View Article and Find Full Text PDFSmall Methods
January 2025
Division of Molecular and Medical Genetics, Center for Gene and Cell Therapy, The Institute of Medical Science, The University of Tokyo, Tokyo, 108-8639, Japan.
Achieving safe and efficient gene therapy hinges upon the inspection of genomes enclosed within individual nano-carriers to mitigate potential health risks associated with empty or fragment-filled vectors. Here solid-state nanopore sensing is reported for identifications of intermediate adeno-associated virus (AAV) vectors in liquid. The method exploits the phenomenon of translocation slowdown induced by the viscosity of salt water-organic mixtures.
View Article and Find Full Text PDFHum Gene Ther
December 2024
Prevail Therapeutics, New York, New York, USA.
Recombinant adeno-associated virus (AAV) is one of the main viral vector-based gene therapy platforms. AAV is a virus consisting of a ≈25 nm diameter capsid with a ≈4.7 kb cargo capacity.
View Article and Find Full Text PDFAnat Histol Embryol
January 2025
National Research System and PEDECIBA, Montevideo, Uruguay.
The Bardhoka sheep breed, also known as Dukagjini white sheep (Ovis aries), is considered to be a medium-sized sheep that is a typical grazing ruminant. The aim of this study was to investigate and measure the parameters of the gastrointestinal tract (GIT) of Bardhoka sheep to determine the morphometry, volume, ratio and surface areas of its different parts. The GIT of eight adult healthy Bardhoka sheep with an average body mass (BM) of 57.
View Article and Find Full Text PDFPlant Cell Rep
December 2024
Institute of Nanfan & Seed Industry, Guangdong Academy of Sciences, Guangzhou, 510000, Guangdong, China.
A total of 24 genes of vacuolar H-translocating pyrophosphatases H-PPases (VPP) genes were identified in Saccharum spontaneum AP85-441 and the ScVPP1-overexpressed Arabidopsis plants conferred salt tolerance. The vital role of vacuolar H-translocating pyrophosphatases H-PPases (VPP) genes involved in plants in response to abiotic stresses. However, the understanding of VPP functions in sugarcane remained unclear.
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