Preparation and in vitro evaluation of an erythrocyte-based butyrylcholinesterase delivery system
10.7644/j.issn.1674-9960.2025.06.009
- VernacularTitle:红细胞载丁酰胆碱酯酶给药系统的制备及体外评价
- Author:
Zhe WANG
1
;
Changwen NING
;
Huaying AN
;
Xingwei JIANG
;
Jun MA
;
Fenghua GAO
;
Pengyu LIU
;
Yanan SUN
;
Ru LI
;
Jinlong LI
;
Yuanyuan YUAN
;
Qun YU
Author Information
1. 军事科学院军事医学研究院,血液安全保障技术研究北京市重点实验室,北京 100850
- Keywords:
organophosphorus nerve agents;
drug-loaded erythrocytes;
butyrylcholinesterase;
hypotonic preswelling method
- From:
Military Medical Sciences
2025;49(6):458-464
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop an erythrocyte-based delivery system for butyrylcholinesterase(BChE)that is capable of prophylaxis against organophosphorus nerve agents.Methods Recombinant BChE was produced and analyzed for oligomerization via polyacrylamide gel electrophoresis(PAGE)and Western blotting.A modified hypotonic preswelling method was employed to prepare BChE-loaded erythrocytes.The drug loading capacity and encapsulation efficiency were quantified using enzyme-linked immunosorbent assay(ELISA).Catalytic activity was assessed in vitro with an activity detection kit.The system was characterized via scanning electron microscopy(SEM),flow cytometry and a hematology analyzer.Results Recombinant BChE predominantly existed as dimers(85％dimer,15％monomer).The optimized volume ratio of erythrocytes to hypotonic solution was determined as 1:7.Compared with native and empty erythrocytes,BChE-loaded erythrocytes exhibited significantly higher catalytic activity(P＜0.001).The mean corpuscular volume of BChE-loaded erythrocytes increased(P＜0.001),while the mean content of corpuscular hemoglobin and hemoglobin in erythrocytes per 100 mL decreased(P＜0.001).SEM revealed no morphological differences(biconcave disc shape).Hypotonic preswelling moderately increased erythrocyte apoptosis(P＜0.001),but no statistical difference was observed between BChE-loaded and hypotonic-treated erythrocytes(P＞0.05).CD47 expression remained unchanged compared to native erythrocytes(P＞0.05).Conclusion The modified hypotonic preswelling method can generate BChE-loaded erythrocytes that retain the characteristics of native erythrocytes while conferring catalytic activity,offering a novel strategy for clinical intervention against organophosphorus poisoning.
