Presentation Information
[P04-608]Poly(β-amino ester) polymer library with monomer variation for mRNA delivery
○Won Jong Kim Kim1, Han Soo Park2, Il Keun Kwon3 (1. Pohang University of Science and Technology (Korea), 2. School of Integrative Engineering, Chung-Ang University (Korea), 3. Department of Dental Materials, School of Dentistry, Kyung Hee University (Korea))
Keywords:
mRNA,Polymer
[Purpose]
Poly(β-amino ester) (PBAE) is a polymer synthesized through the polymerization of amines and acrylates. By varying the amine and acrylate monomers, PBAEs can adopt diverse structures, allowing for the synthesis of polymers tailored for mRNA delivery through the combination of different monomers. The synthesized polymers are biodegradable, resulting in low toxicity after injection. Additionally, the structure of the monomers influences differences in mRNA delivery efficiency and immune activation.
[Method]
Polymers demonstrating high mRNA expression efficiency were selected and complexed with 6 μg of Covid-19 mRNA, followed by intramuscular injection into mice. Two weeks after injection, serum was collected to assess neutralizing antibody formation using enzyme-linked immunosorbent assay (ELISA) and to evaluate viral neutralization capacity using a plaque reduction neutralization test (PRNT). Additionally, splenocytes were harvested, and interferon-γ production was measured via enzyme-linked immunospot (ELISpot) assays to confirm T cell activation.
[Results]
Seven polymers with high mRNA expression efficiency were selected for further experiments. As a positive control, Lipid nanoparticles (LNP) formulated with SM-102 was used. Serum samples were collected and diluted at 2x, 5x, 10x, 20x, and 40x for ELISA. The antibody levels in the blood were lower for polymers compared to LNP (Figure 1A). However, when viral neutralization against the COVID-19 virus (Wuhan strain) was evaluated using a plaque reduction neutralization test (PRNT), polymer 8D exhibited significant viral neutralization activity (Figure 1B). This suggests that although the total amount of antibodies against the spike protein was lower, the proportion of neutralizing antibodies was higher.Additionally, splenocytes were harvested, treated with the spike protein, and analyzed using IFN-γ ELISpot, which showed that polymer 8B induced high levels of IFN-γ production (Figure 1C). These results demonstrate that polymer 8D promotes the formation of neutralizing antibodies, while polymer 8B induces T cell immune activation, confirming the potential of polymer-based mRNA delivery
[Consideration]
[Conclusion]
This study suggests that immune responses can be induced not only by lipid nanoparticles but also by delivering mRNA using polymers. Further optimization of polymer length, particle size, and the ratio of polymer to mRNA could enable the development of mRNA vaccines utilizing polymers.
Poly(β-amino ester) (PBAE) is a polymer synthesized through the polymerization of amines and acrylates. By varying the amine and acrylate monomers, PBAEs can adopt diverse structures, allowing for the synthesis of polymers tailored for mRNA delivery through the combination of different monomers. The synthesized polymers are biodegradable, resulting in low toxicity after injection. Additionally, the structure of the monomers influences differences in mRNA delivery efficiency and immune activation.
[Method]
Polymers demonstrating high mRNA expression efficiency were selected and complexed with 6 μg of Covid-19 mRNA, followed by intramuscular injection into mice. Two weeks after injection, serum was collected to assess neutralizing antibody formation using enzyme-linked immunosorbent assay (ELISA) and to evaluate viral neutralization capacity using a plaque reduction neutralization test (PRNT). Additionally, splenocytes were harvested, and interferon-γ production was measured via enzyme-linked immunospot (ELISpot) assays to confirm T cell activation.
[Results]
Seven polymers with high mRNA expression efficiency were selected for further experiments. As a positive control, Lipid nanoparticles (LNP) formulated with SM-102 was used. Serum samples were collected and diluted at 2x, 5x, 10x, 20x, and 40x for ELISA. The antibody levels in the blood were lower for polymers compared to LNP (Figure 1A). However, when viral neutralization against the COVID-19 virus (Wuhan strain) was evaluated using a plaque reduction neutralization test (PRNT), polymer 8D exhibited significant viral neutralization activity (Figure 1B). This suggests that although the total amount of antibodies against the spike protein was lower, the proportion of neutralizing antibodies was higher.Additionally, splenocytes were harvested, treated with the spike protein, and analyzed using IFN-γ ELISpot, which showed that polymer 8B induced high levels of IFN-γ production (Figure 1C). These results demonstrate that polymer 8D promotes the formation of neutralizing antibodies, while polymer 8B induces T cell immune activation, confirming the potential of polymer-based mRNA delivery
[Consideration]
[Conclusion]
This study suggests that immune responses can be induced not only by lipid nanoparticles but also by delivering mRNA using polymers. Further optimization of polymer length, particle size, and the ratio of polymer to mRNA could enable the development of mRNA vaccines utilizing polymers.
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