Targeting of M13 Phage Particles by Human Holotransferrin Using a Chemical Coupling Strategy

Abstract

Background & Objectives: Targeting transgene carriers and vectors to individual cells and tissues is one of the most important goals of gene therapy. Bacteriophages are of appropriate transgene carriers and there are different methods for their targeting to target cells. Present study reports preparation of targeted M13-based bacteriophage particles by a chemical coupling strategy. Materials and Methods: First, the pCMV-Script-GFP construct was produced via in vivo excision protocol fromGFP Phage particles using ExAssist helper phage and XLOLR as specific host. Then, M13 phage particles bearing GFP (M13-GFP) were obtained by single stranded rescue using R408 helper phage. The human holotransferrin molecules were then coupled to the surface of phage particles by reductive amination chemistry. Transferrin molecules bind to the surface of phage particles were studied by phage-ELISA. Results: Phage-ELISA tests showed that holotransferrin molecules were coupled to the surface of M13 phage particles in a correct way and the transferrin-targgeted M13 phage particles were prepared. Further analysis showed that about 485 transferrin molecules coupled per phage particle. Conclusion: The results show that chemical coupling might be considered as a suitable strategy for targeting of M13 particles via coupling of targeting molecules in high density to the phage surface.