Cytocompatibility of protein nanocage on mammalian cell line

• Main Author: May, Moe Thu
• Other Authors: Lim Sierin
• Format: Final Year Project
• Language: English
• Published: 2015
• Subjects: DRNTU::Engineering::Bioengineering
• Online Access: http://hdl.handle.net/10356/65090
• Institution: Nanyang Technological University

Archaeoglobus fulgidus ferritin mutant, AfFtn-AA has been shown to be a potential MRI contrast agent and therapeutic carriers. However, the reactivity and host immunological response is not yet established. In this study, cytocompatibility of AfFtn- AA and iron loaded AfFtn-AA were evaluated by measuring the release of reactive oxygen species (ROS) produced by inflammatory cells. Murine macrophages, Raw 264.7 cells, were used to conduct the study as they are known to produce ROS when exposed to foreign materials. ROS is detected by luminol assay and conditions for optimal detection of ROS were investigated under a variety of conditions. ROS production was determined by varying cell concentrations, ROS stimulation with lipopolysaccharide (LPS) and Phorbol 12-Myristate 13-Acetate (PMA). Using luminol as a probe, different levels of ROS were observed for 0.1mg/ml and 0.01mg/ml AfFtn-AA and iron loaded AfFtn-AA at different timepoint. These data are the first in vitro demonstration of ROS generation in response to AfFtn-AA, and luminol describes a novel technique to evaluate the host response.