Interviewee
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Bobo DANG
School of Life Sciences
Biology
Laboratory of Protein Engineering and Protein Therapeutics
Looking for
We contacted him due to his extensive expertise in protein design and biopresentation technology, and we are seeking his guidance on the feasibility of implementing protein-coupled nanoparticles in PEG and achieving direct conjugation in prokaryotic organisms.
Question we asked
- When isolating and synthesizing iron oxide nanoparticles from E. coli, we encountered the need to remove a significant amount of biomaterial. Could you please recommend an effective washing agent that can achieve good removal results without significantly damaging the surface composition of the iron oxide nanoparticles?
- How can we achieve the coupling between antibodies and the surface of iron oxide nanoparticles? We are considering two approaches: direct coupling to the isolated iron oxide nanoparticles, where the surface coating component may be phospholipids, and pre-coating the nanoparticles with a layer of PEG (polyethylene glycol) before coupling. Do you have any suggestions for the design of coupling components? We have thought about the Spytag/Spycatcher system.
- Is it possible to enhance the connection between the tail end of the antibodies and the nanoparticles through a biocompatible method of modifying the binding site?
- After synthesizing our antibodies, we aim to improve their targeting efficiency. Are there any biocompatible methods available to modify the binding site at the head end of the antibodies?
Takeaways
- During our discussion on the self-assembly of bio-synthetic nanoparticles and antibodies in Escherichia coli, professor Dang expressed concerns regarding the stability of the vesicles on the surface of the nanoparticles compared to the bacterial membrane. Professor Dang suggested that this instability might lead to the destruction of the nanoparticle structure when separating them with antibodies. Consequently, he proposed that it would be more feasible for us to synthesize and assemble the components separately.
- Another topic of our discussion was elution during the separation of nanoparticles. professor Dang suggested that we could employ organic solvents for washing and filtration. Additionally, considering the necessity of a layer of PEG membrane, professor Dang recommended attempting to dissolve PEG in organic solvents to achieve the desired replacement.
- Professor Dang opined that the surface composition of the nanoparticles is not necessarily crucial. As long as the final objective of antibody connectivity is achieved, the existence of amino and carboxyl groups on the surface of bio-synthesized nanoparticles should facilitate connection. Professor Dang also proposed that the possibility of employing harsh solvents to wash the nanoparticles and observe their binding behavior with antibodies.
- In relation to connecting the antibodies and nanoparticles, professor Dang mentioned the utilization of EDC (1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide).
- Overall, an additional point to consider is that using PEG coating might not be meaningful. Commercially available chemically-synthesized nanoparticles are already coated with PEG. Therefore, the coating on the surface of bio-synthesized nanoparticles lacks significance.