His-Tag-Aptamer-Liposome-Conjugates as a Multipurpose Tool for Recombinant Protein Screening and Protein-Based...

Area of Science:

• Bioconjugation Chemistry
• Nanotechnology in Bioanalysis
• Molecular Recognition

Background:

• Liposome-aptamer conjugates combine aptamer specificity with liposome encapsulation for bioanalysis and drug delivery.
• His-tags are widely used for protein purification and detection, but current methods can be labor-intensive and costly.

Purpose of the Study:

• To develop multipurpose liposome-aptamer conjugates for specific binding to his-tags.
• To evaluate these conjugates as screening tools for his-tagged proteins and signaling tools in bioassays.
• To optimize aptamer immobilization on liposomes while maintaining aptamer functionality and liposome integrity.

Main Methods:

• Synthesized liposome-aptamer conjugates using various immobilization strategies: cholesterol-functionalized, amine-modified, and biotinylated aptamers.
• Characterized liposome and aptamer properties and conjugate functionality using heterogeneous binding assays.
• Assessed the stability of the conjugates at 4 °C for at least 4 months.

Main Results:

• Post-insertion and biotinylated aptamer immobilization methods yielded stable and functional his-aptamer-liposome conjugates.
• These conjugates demonstrated stability for at least 4 months at 4 °C.
• Other immobilization methods negatively impacted aptamer structure and liposome integrity.

Conclusions:

• Developed a stable and versatile platform technology using his-tag aptamer-liposome conjugates.
• These conjugates can be used for efficient screening of his-tagged proteins and as signaling tools in bioassays.
• The platform offers improved assay control, reproducibility, and cost-effectiveness compared to traditional methods.