2020 Fiscal Year Research-status Report
Design of degradable glycopolymeric nanoparticles for the safe and targeted delivery of proteins
| Project/Area Number |
20K20197
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| Research Institution | Japan Advanced Institute of Science and Technology |
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Principal Investigator |
Rajan Robin 北陸先端科学技術大学院大学, 先端科学技術研究科, 助教 (70848043)
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| Project Period (FY) |
2020-04-01 – 2023-03-31
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| Keywords | Micelles / Protein / Zwitterionic polymer / Degradable / Glycopolymer |
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| Outline of Annual Research Achievements |
This year, I prepared glycopolymer-based micelles for the protection of proteins against stress. I selected Trehalose as the sugar unit for the initial study because of its well-known protein protecting property. The synthesis of the micelle was done in three steps. In the first step, trehalose was functionalized with the methacrylate group. At the same time, ring-opening polymerization of caprolactone was performed using a hydroxyl group-containing RAFT agent, which yielded polycaprolactone macro-chain transfer agent (PCL-CTA). In the final step, trehalose methacrylate, sulfobetaine monomer (which has excellent protein protecting property) were polymerized in the presence of PCL-CTA to get degradable micelles.
For the analysis of the protein stabilization, I chose Lactate dehydrogenase (LDH). The thermal aggregation of LDH was completely inhibited by these micelles at very low concentrations (>1 mg/mL). Activity of LDH was also retained when incubated in the presence of these micelles.
Most importantly, the micelles could be easily separated from the protein by ultracentrifugation, which has been the biggest shortcoming with the polymer based inhibitors of protein aggregation. The isolated protein was recovered with all its activity retained.
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| Current Status of Research Progress |
Current Status of Research Progress
1: Research has progressed more than it was originally planned.
Reason
The plan for this year was to develop and optimize the conditions for the development of zwitterionic glycopolymeric micelles, which I have successfully managed to do. In addition, I have been able to study the protein stabilizing property with the micelles, which yielded excellent results. Further, I managed to develop a system to isolate the protein from the stabilizing agent.
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| Strategy for Future Research Activity |
In the coming months, I will perform a detailed analysis of the protein protection property of the micelles. For this, I will use different proteins such as lysozyme, insulin, etc. At the same time, I will check its activity in protecting relevant antibodies and enzymes. This will be followed by studying the stability and degradability of these micelles under different conditions.
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| Causes of Carryover |
This year involved only polymer synthesis and its characterization, hence a lot of budget was not required. however, in the next fiscal year, I plan to test the activity of micelles against various proteins and enzymes. hence, more money would be needed.
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