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Evaluation of interactions between RAW264.7 macrophages and small molecules by capillary electrophoresis

Wang, Feng‐Qin, Li, Qiao‐Qiao, Zhang, Qian, Wang, Yin‐Zhen, Hu, Yuan‐Jia, Li, Peng, Wan, Jian‐Bo, Yang, Feng‐Qing, Xia, Zhi‐Ning
Electrophoresis 2017 v.38 no.6 pp. 938-941
capillary electrophoresis, chemical elements, electric field, free air carbon dioxide enrichment, macrophages, naringin, oleuropein, stoichiometry, viscosity
In this study, the affinity interactions between RAW 264.7 macrophages and three small molecules including naringin, oleuropein and paeoniflorin were evaluated by affinity capillary electrophoresis (ACE), partial filling affinity capillary electrophoresis (PFACE) and frontal analysis capillary electrophoresis (FACE), respectively. The result indicated that ACE (varying concentrations of cell suspension were filled in the capillary as receptor) may not be suitable for the evaluation of interactions between cell and small molecules due to the high viscosity of cell suspension; PFACE can qualitatively evaluate the interaction, but the difference in viscosity between RAW264.7 suspension and buffer effects on the liner relationship between filling length and injection time, which makes the calculation of binding constant difficult. Furthermore, based on the PFACE results, naringin showed stronger interaction with macrophages than the other two molecules; taking advantage of the aggregation phenomenon of cell induced by electric field, FACE was successfully used to determine the stoichiometry (n = 5×10⁹) and binding constant (Kb = 1×10⁴ L/mol) of the interaction between RAW264.7 and naringin.