PXD070195

PXD070195 is an original dataset announced via ProteomeXchange.

Title	Scalable purification enables high-quality virus-like particles for therapeutic translation
Description	Emerging molecular therapies introduce enzymatic activity into cells by delivering genes, transcripts, or proteins. Owing to their robust cell-entry capacity, virus-like particles (VLPs) represent a technology of choice in genome editing, where low doses of heterologous proteins and nucleic acids are essential. However, clinical translation of VLP vectors is hindered by inadequate purification methods. Current approaches, relying primarily on ultracentrifugation, suffer from inconsistent product quality and poor scalability. Here, we report the development of a broadly applicable purification strategy that improves the purity and therapeutic efficacy of genome-editing VLPs. Considering the characteristic properties of murine leukemia virus (MLV)-derived engineered VLPs (eVLPs) and HIV-derived engineered nucleocytosolic vehicles for loading of programmable editors (ENVLPEs+), we developed a workflow that involves single- and multi-modal chromatographic steps, effectively removing host cell proteins and cell-culture contaminants while improving VLP integrity and biological activity. Our purified VLPs displayed superior protein composition, consistency, and enhanced functional delivery compared to VLPs partially purified by conventional ultracentrifugation methods. Mass spectrometric analysis revealed a substantial decrease in contaminants, with VLP-specific proteins comprising >90% of the final product. In vivo studies confirmed improved therapeutic outcomes when chromatographically purified VLPs were used. Our scalable purification platform addresses critical manufacturing bottlenecks and constitutes a starting point for further development of VLP therapeutics, enabling robust production of pure VLPs for diverse applications such as genome editing, vaccine development, and other uses that require intracellular protein delivery.
HostingRepository	PRIDE
AnnounceDate	2025-11-26
AnnouncementXML	Submission_2025-11-26_00:01:25.283.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Fangyuan Gao
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2025-11-02 09:13:07	ID requested
⏵ 1	2025-11-26 00:01:26	announced

Ho, ł, ubowicz R, Gao F, Du SW, Rodrigues Menezes C, Zhang J, Ho, ł, ubowicz MW, Chen PZ, Armbrust N, Geilenkeuser J, Liu DR, Truong DJ, Westmeyer GG, Palczewska G, Palczewski K, Scalable purification enables high-quality virus-like particles for therapeutic translation. J Biol Chem, 301(12):110946(2025) [pubmed]

10.1016/j.jbc.2025.110946;

submitter keyword: , prime editing, protein delivery, CRISPR/Cas, nanotechnology, Cas9,chromatography, virus-like particles, base editing, vesicles, ribonuclear protein (RNP)

Krzysztof Palczewski
contact affiliation	1Gavin Herbert Eye Institute –Brunson Center for Translational Vision Research, Department of Ophthalmology and Visual Sciences, University of California Irvine, Irvine, CA 92697, USA 2Department of Physiology and Biophysics, University of California Irvine, Irvine, CA 92697, USA 3Department of Chemistry, University of California Irvine, Irvine, CA 92697, USA 4Department of Molecular Biology and Biochemistry, University of California Irvine, Irvine, CA 92697, USA
contact email	kpalczew@uci.edu
lab head
Fangyuan Gao
contact affiliation	University of California, Irvine
contact email	fangyuag@hs.uci.edu
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2025/11/PXD070195

PRIDE project URI

• PRIDE
  1. PXD070195
     1. Label: PRIDE project
     2. Name: Scalable purification enables high-quality virus-like particles for therapeutic translation