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

Abstract

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 = 5×10 ) and binding constant (K = 1×10 L/mol) of the interaction between RAW264.7 and naringin.

Studying molecular interactions via capillary electrophoresis and microscale thermophoresis: A review

“…Most often, we encounter the determination of binding parameters between proteins and drugs [47,52,57,[145][146][147][148]. However, CE-FA was used to characterize the interactions of glycosaminoglycans-protein [149], macrophage-drug [150], heme-insulin [151], platelets-small molecules [152], CD-small molecules [153], ss-DNA-small molecules [49], antigen-adjuvant [58], polymer-copolymer [59,60,154], and protein-polysaccharide [55].…”

“…It has been found a strong dependence of the binding parameters with the ionic strength and also that the concentration of polymeric adjuvant significantly modifies the ionic strength of the formulation, the extent of which can be estimated and corrected [58]. The applications of CE-FA [47,52,55,57,[145][146][147][148][149][150][151][152][153][155][156][157][158][159][160][161] and FACCE [58][59][60]154] are summarized in Table 3.…”

Capillary electrophoresis in phytochemical analysis (2014–2017)

“…Results indicated that ACE may not be suitable for the evaluation of interactions between cell and small molecules due to the high viscosity of cell suspension, and in PFACE the difference in viscosity between cell suspension and buffer effects on the linear relationship between filling length and injection time, which makes the calculation of binding constant difficult. FACE was successfully used to determine the stoichiometry and binding constant of the interaction between RAW264.7 and naringin, by taking advantage of the aggregation phenomenon of cell induced by electric field [133]. Platelets play a vital role in some physical processes as homeostasis, thrombosis and inflammation.…”