Rapid, Scalable, and Cost-Effective Manufacturing of Uniform Non-Enveloped, Tag-Free Virus-Like Particles.

Virus-like particles (VLPs) are widely recognized as safe and versatile nanostructures with broad applications in vaccine development, gene delivery, and nanotechnology, but their production typically requires time-consuming procedures, incurs high costs, and yields products of limited purity. This article outlines a rapid, scalable, and cost-effective method for producing high-quality VLPs using the VP1s capsid protein from murine polyomavirus. The procedure comprises four principal stages: expression of the capsid protein in Escherichia coli, extraction and stabilization of the protein, purification to yield high-quality capsomeres, and controlled in vitro assembly of VLPs. Compared with conventional protocols that rely on in vivo assembly and labor-intensive purification procedures, such as ultracentrifugation- or affinity tag-based methods, the described approach provides a streamlined and rapid workflow that avoids affinity tags, achieves consistently high yields, and enables precise regulation of assembly conditions, resulting in a scalable and highly cost-efficient strategy for VLP production within 5 days. © 2026 Wiley Periodicals LLC. Basic Protocol 1: Expression and preparation of polyomavirus VP1 protein in E. coli Basic Protocol 2: Polymomavirus VP1 purification Basic Protocol 3: VP1 assembly Basic Protocol 4: Quality control.