Construction and Functionalization of a Clathrin Assembly for a Targeted Protein Delivery

• Published in: Small (Weinheim an der Bergstrasse, Germany) Vol. 19; no. 8; pp. e2204620 - n/a
• Main Authors: Kim, Hong‐Sik, Bae, Jin‐Ho, Kim, Gijeong, Song, Ji‐Joon, Kim, Hak‐Sung
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 01.02.2023
• Subjects: Assemblies; Assembly; avidity; Biocompatibility; Biomedical materials; Cargo; Chains; clathrin; Clathrin - metabolism; Drug Delivery Systems; functionalization; Humans; Nanotechnology; protein assembly; protein delivery; Proteins; avidity; functionalization; clathrin; protein delivery; protein assembly
• ISSN: 1613-6810; 1613-6829; 1613-6829
• DOI: 10.1002/smll.202204620

Protein assemblies have drawn much attention as platforms for biomedical applications, including gene/drug delivery and vaccine, due to biocompatibility and functional diversity. Here, the construction and functionalization of a protein assembly composed of human clathrin heavy chain and light chain for a targeted protein delivery, is presented. The clathrin heavy and light chains are redesigned and associated with each other, and the resulting triskelion unit further self‐assembled into a clathrin assembly with the size of about 28 nm in diameter. The clathrin assembly is dual‐functionalized with a protein cargo and a targeting moiety using two different orthogonal protein–ligand pairs through one‐pot reaction. The functionalized clathrin assembly exhibits about a 900‐fold decreased KD value for a cell‐surface target due to avidity compared to a native targeting moiety. The utility of the clathrin assembly is demonstrated by an efficient delivery of a protein cargo into tumor cells in a target‐specific manner, resulting in a strong cytotoxic effect. The present approach can be used in the creation of protein assemblies with multimodality. A new type of protein self‐assembly using human clathrin is developed. Using two different orthogonal protein–ligand pairs, both heavy and light chains of a clathrin triskelion are redesigned for a dual‐functionalization with functional biomolecules. The resulting dual‐functionalized clathrin assembly shows a target‐specific protein delivery and exemplifies its utility as a protein delivery platform.