Key Laboratory of Drug Quality Control and Pharmacovigilance, Ministry of Education , China Pharmaceutical University , Nanjing 210009 , China.
Department of Biochemistry, School of Life Science and Technology , China Pharmaceutical University , Nanjing 210009 , China.
ACS Sens. 2019 Apr 26;4(4):977-983. doi: 10.1021/acssensors.9b00102. Epub 2019 Mar 21.
The high-throughput screening of chemically active substances has aroused widespread interest in recent years. The screening of drug carriers is usually ignored, although they interact directly with physiological barriers and target cells, and they determine the fate and efficacy of drugs in vivo. In this work, a series of polydiacetylene (PDA) vesicles (ca. 550 nm) that simulate the cell membrane are constructed to detect the membrane affinity of gene vectors. The surface potentials of vesicles are adjusted by changing the phospholipid composition using different charged compounds. All vesicles show the rapid color changes upon the addition of gene vectors by the naked eye within <5 min. The optimized 1,2-dimyristoyl- sn-glycero-3-phosphocholine (DMPC)-PDA vesicles display the most sensitive discoloration response to the commercially available gene vectors, including Lipofectamine 2000, Entranster-H4000, and polyethylenimine. The logarithm of transfection efficiency for green fluorescent protein plasmid (pGFP) mediated by these three vectors in L02 and HepG2 cells demonstrate an excellent linear correlation with the logarithm of membrane affinity (log K) of the gene vectors detected by DMPC-PDA vesicles. This rapid visualization method not only allows the in vitro membrane affinity prediction of gene vectors that greatly contributes to the gene transfection efficiency, but also offers a universal strategy for the potential high-throughput screening of various carrier materials featuring high cell affinity.
近年来，高通量筛选化学活性物质引起了广泛的关注。然而，药物载体的筛选通常被忽视，尽管它们直接与生理屏障和靶细胞相互作用，并且决定了药物在体内的命运和疗效。在这项工作中，构建了一系列模拟细胞膜的聚二乙炔（PDA）囊泡（约 550nm），以检测基因载体的膜亲和力。通过使用不同带电化合物改变磷脂组成来调整囊泡的表面电势。所有囊泡在加入基因载体后通过肉眼在<5 分钟内迅速发生颜色变化。优化后的 1,2-二肉豆蔻酰基-sn-甘油-3-磷酸胆碱（DMPC）-PDA 囊泡对市售基因载体（包括 Lipofectamine 2000、Entranster-H4000 和聚乙烯亚胺）显示出最敏感的变色响应。这三种载体介导的绿色荧光蛋白质粒（pGFP）的转染效率的对数与通过 DMPC-PDA 囊泡检测到的基因载体的膜亲和力（log K）的对数呈极好的线性相关。这种快速可视化方法不仅可以预测基因载体的体外膜亲和力，这对基因转染效率有很大贡献，而且还为具有高细胞亲和力的各种载体材料的高通量筛选提供了一种通用策略。
