A new biochromatography model based on DNA origami assembled PPARγ: construction and evaluation

Area of Science:

• Biochemistry
• Nanotechnology
• Analytical Chemistry

Background:

• Receptors are valuable drug targets, but immobilizing them on supports like silica often compromises their bioactivity due to poor biocompatibility.
• DNA origami offers excellent biocompatibility and can serve as a scaffold to improve receptor assembly and silica carrier performance.

Purpose of the Study:

• To construct a novel biochromatography model utilizing DNA origami for enhanced receptor immobilization and drug screening.
• To improve the biocompatibility of silica supports for receptor-based drug discovery.

Main Methods:

• Self-production of M13 single-stranded DNA (ssDNA) for cost-effective scaffolding.
• Assembly of the ligand-binding domain of peroxisome proliferator-activated receptor gamma (PPARγ-LBD) onto DNA origami carriers.
• Coupling of DNA origami-PPARγ-LBD constructs onto silica surfaces to create a stationary phase for biochromatography.
• High-performance liquid chromatography (HPLC) was used to evaluate the affinity and specificity of the developed biochromatography model.

Main Results:

• The constructed biochromatography model demonstrated specific recognition of rosiglitazone, indicating its ability to screen chemical compounds interacting with PPARγ.
• This represents the first use of DNA origami to assemble PPARγ for biochromatography construction.
• The method proved efficient, convenient, and high-throughput for drug screening applications.

Conclusions:

• The DNA origami-based biochromatography model offers a significant advancement in drug target screening.
• This innovative approach provides a new, rapid, and convenient method for screening active ingredients from complex natural product extracts and libraries.
• The enhanced biocompatibility and specific recognition capabilities pave the way for broader applications in pharmaceutical research and development.