Scalable purification enables high-quality virus-like particles for therapeutic translation

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 ce..., , # Scalable purification enables high-quality virus-like particles for therapeutic translation

## Description of the data and file structure

The data is organized in compressed subdirectories whose names correspond to figures in the main directory (Figure_1, Figure_2, Figure_S1, etc.). Exception: Figure S3 is the uncropped, unadjusted image of the gel.

Figure_1.zip

Figure_1a: Anti-Cre immunoblot of Cre-eVLP before and after purification by sucrose cushion ultracentrifugation. Rafal H 2024-02-25 20h25m53s(Chemiluminescence).jpg = Chemiluminescence scan of the immunoblot. Rafal H 2024-02-25 20h25m53s(Cy3) = red channel scan of the immunoblot. Rafal H 2024-02-25 20h25m53s(Cy5).jpg = blue channel scan of the immunoblot.

Figure1b_VLP!_170713-2.jpg: A representative electron microscopic image of purified eVLPs.

Figure_1c,d: Quantification of mCherry delivery into HEK 293T cells by fluorescence microscopy. Brightfield = brightfield images of the cells. RFP = red fluorescence images of the...,