Functionalization of cell surface membranes via coating with M13 bacteriophage

Abstract

Cell surface functionalization holds significant potential in various biomedical applications. This study focused on the engineering and characterization of M13 bacteriophage for effective cell surface functionalization. The p8 coat protein of the M13 phage was genetically engineered to introduce four negatively charged glutamates, so that the surface charge of the phage was more negative than that of the wild type. Characterization of the phage was performed by MALDI-TOF, determining the number of maleimides per phage and optimizing the sulfo-SMCC concentration. Chemical modification enabled the exposure of maleimide groups on the phage surface, facilitating their attachment to the reduced cell surface membrane. Confocal fluorescent imaging confirmed the uniform distribution and attachment of the 4E phage onto target cells without detachment or internalization. The functionalized cells exhibited an increased negative surface charge and showed no significant cytotoxicity or reactive oxygen species (ROS) production in T-MSCs and Fadu cells. These results demonstrate the potential of 4E phage as an efficient and cell-friendly tool for cell surface functionalization. However, pro-inflammatory cytokine gene expression levels were found to be elevated, indicating the need for further research and purification processes to optimize its applications in cell-based therapies and biomedical engineering.