H-ferritin nanocages: refined strategy for optimized protein production and purification

H-ferritin (HFn) nanocages are promising tools in biomedical applications, yet producing sufficient injectable quantities remains challenging. To optimize production, we compared two lysis methods and purification protocols, focusing on the ClearColi BL21(DE3) strain, which features a modified lipopolysaccharide that avoids triggering endotoxin responses in humans. Our findings show that omitting protease inhibitors during lysis and using 20 mM Tris-HCl pH 8.0 buffer with Q-Sepharose resin significantly enhances protein yield. Importantly, these adjustments do not compromise encapsulation efficiency or cytotoxicity, offering a more efficient and cost-effective strategy for obtaining pure, stable HFn suitable for therapeutic use.