2. Academic Validation
  3. Recombinant ApoE3 corona plus sucrose vitrification endows mRNA-LNPs with Freeze-Thaw and -80 °C stability

Recombinant ApoE3 corona plus sucrose vitrification endows mRNA-LNPs with Freeze-Thaw and -80 °C stability

  • Int J Pharm. 2025 Nov 30:685:126281. doi: 10.1016/j.ijpharm.2025.126281.
Sai-Ning Tian 1,  Juan-Juan Zhao 2,  Zu-Yi Peng 1,  Jia-Xin Ren 2,  Xi-Die Zhao 2,  Tao Cheng 3,  Jian-Ping Zhang 4,  Xiao-Bing Zhang 5
Affiliations

Affiliations

  • 1 Haihe Laboratory of Cell Ecosystem, Tianjin Medical University, Tianjin, China.
  • 2 State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China; Tianjin Institutes of Health Science, Tianjin 301600, China.
  • 3 State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China; Tianjin Institutes of Health Science, Tianjin 301600, China. Electronic address: [email protected].
  • 4 State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China; Tianjin Institutes of Health Science, Tianjin 301600, China. Electronic address: [email protected].
  • 5 Haihe Laboratory of Cell Ecosystem, Tianjin Medical University, Tianjin, China; State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China; Tianjin Institutes of Health Science, Tianjin 301600, China. Electronic address: [email protected].
PMID: 41101606 DOI: 10.1016/j.ijpharm.2025.126281
Abstract

Lipid nanoparticles (LNPs) enable clinical mRNA delivery but remain paradoxically fragile: storage at -80 °C slows RNA hydrolysis yet triggers lipid phase separation and ice damage. Conventional sugar cryoprotectants immobilize water, but incompletely stabilize the lipid matrix, leaving ＞30 % activity loss after one month. Here we report a dual, chemically-defined strategy that (i) pre-adsorbs recombinant Apolipoprotein E3 (ApoE3) to which may help stabilize lipid packing via reversible interfacial binding, and (ii) vitrifies the surrounding solution with 10 % (w/v) sucrose. Using two representative ALC-0315 backbones with divergent potencies, the dual strategy preserved > 90 % mRNA encapsulation, size uniformity and luciferase expression after 4 weeks at -80 °C and three freeze-thaw cycles. In C57BL/6 mice, liver bioluminescence rose ∼ 3-fold relative to fresh untreated LNPs without elevating interleukin-6(IL-6) or organ toxicity. Thawed samples retained > 70 % potency for 72 h at 4-25 °C, exceeding the ≤ 12 h handling window of authorized COVID-19 vaccines. Both excipients can be produced at cGMP scale and are added in a single step, facilitating integration into existing manufacturing and cold-chain workflows. By combining pre-adsorbed ApoE3 with sucrose vitrification, this strategy provides a practical route to more durable mRNA vaccines and gene therapies while avoiding lyophilization or plasma-derived additives.

Keywords

Apolipoprotein E3; Cryostability; Lipid nanoparticle; Protein corona; mRNA delivery.

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