Construction and optimization of a novel plasma membrane model for membrane affinity assay based on polydiacetylene vesicles
10.16438/j.0513-4870.2022-1129
- VernacularTitle:基于聚联乙炔囊泡的新型膜亲和力测定模型的构建和优化
- Author:
Chai-chai NIE
;
Rui-ting DONG
;
Yu-tong WU
;
Jing-bo WU
;
Sheng ZHANG
;
Feng ZHENG
;
Ya DING
- Publication Type:Research Article
- Keywords:
DMPC-PDA vesicle;
membrane affinity;
visualization;
model optimization;
plasma membrane model;
stability
- From:
Acta Pharmaceutica Sinica
2023;58(8):2503-2511
- CountryChina
- Language:Chinese
-
Abstract:
Most drugs need to interact with cell membrane to reach the biological target, so that membrane affinity assay is an important early screening step in drug discovery. However, at present, the traditional oil-water distribution method is still used, a new, simple and accurate method for membrane affinity assay is urgently needed. In this study, according to the colorimetric principle, a new assay model based on polydiacetylene vesicles was optimized through a series of experiments including different concentrations of vesicle solution, temperature, or pH reaction environment. On this basis, tetracaine hydrochloride, 2-methylimidazole and histamine were used as model drugs to measure the membrane affinity constants and verify the between-batch precision of the optimized assay model (relative standard deviation less than 5%). In addition, polydiacetylene vesicles were stable for up to 180 days, demonstrating the potential application of the assay model. This strategy is simple, stable, reliable, with high reproducibility, low cost and easy to promote, which provided a new tool and a new direction for the high-throughput assay of membrane affinity.
