Department of Pharmaceutics and Biopharmaceutics, Johannes Gutenberg University Mainz, D-55131 Mainz, Germany.
BioNTech RNA Pharmaceuticals, D-55131 Mainz, Germany.
Cells. 2020 Sep 5;9(9):2034. doi: 10.3390/cells9092034.
Hybrid nanoparticles from lipidic and polymeric components were assembled to serve as vehicles for the transfection of messenger RNA (mRNA) using different portions of the cationic lipid DOTAP (1,2-Dioleoyl-3-trimethylammonium-propane) and the cationic biopolymer protamine as model systems. Two different sequential assembly approaches in comparison with a direct single-step protocol were applied, and molecular organization in correlation with biological activity of the resulting nanoparticle systems was investigated. Differences in the structure of the nanoparticles were revealed by thorough physicochemical characterization including small angle neutron scattering (SANS), small angle X-ray scattering (SAXS), and cryogenic transmission electron microscopy (cryo-TEM). All hybrid systems, combining lipid and polymer, displayed significantly increased transfection in comparison to lipid/mRNA and polymer/mRNA particles alone. For the hybrid nanoparticles, characteristic differences regarding the internal organization, release characteristics, and activity were determined depending on the assembly route. The systems with the highest transfection efficacy were characterized by a heterogenous internal organization, accompanied by facilitated release. Such a system could be best obtained by the single step protocol, starting with a lipid and polymer mixture for nanoparticle formation.
脂质体和聚合物混合纳米颗粒被组装起来，用作信使 RNA（mRNA）转染的载体，使用不同比例的阳离子脂质 DOTAP（1,2-二油酰基-3-三甲基铵丙烷）和阳离子生物聚合物鱼精蛋白作为模型系统。与直接的单步方案相比，应用了两种不同的顺序组装方法，并研究了与所得纳米颗粒系统的生物活性相关的分子组织。通过包括小角中子散射（SANS）、小角 X 射线散射（SAXS）和低温透射电子显微镜（cryo-TEM）在内的彻底物理化学特性研究，揭示了纳米颗粒的结构差异。与单独的脂质/mRNA 和聚合物/mRNA 颗粒相比，所有结合脂质和聚合物的混合系统的转染效率均显著提高。对于混合纳米颗粒，根据组装路线，确定了内部组织、释放特性和活性的特征差异。转染效率最高的系统具有异质的内部组织，并伴有促进释放。通过从脂质和聚合物混合物开始形成纳米颗粒的单步方案，可以获得这种具有最高转染效率的系统。
