Capillary electrophoresis using immobilized whole cells with overexpressed endothelin receptor for specific ligand screening

Abstract

A new capillary electrophoresis method using immobilized cells as the stationary phase has been developed. The power of this method is demonstrated by the separation and identification of endothelin antagonists on a capillary column coated by the transfected Chinese hamster ovary (CHO) cells with overexpressing endothelin receptors. The screening results are validated by functional assays suppressing the increase of intracellular calcium concentration induced by endothelin-1. Instead of making efforts in isolating protein receptors, the easily prepared whole-cell capillary column provides a superior tool on the basis of ligand/receptor affinity for a rapid screening of potent drug candidates from compound libraries.

CCR4 expressing cells cultured adherently on a capillary wall and formaldehyde fixed as the stationary phase for ligand screening by ACE

“…The screening activity of this capillary was still observed after keeping for 30 days. This is longer than the durability of other kinds of the cell‐immobilized capillaries reported so far .…”

“…However, this method with a complex preparation procedure is also costly and time‐consuming. Owing to the attractive features of CE mentioned above, a couple of new methods were developed for immobilizing whole cells with over expressing receptors on the inner wall of capillary column in ACE . Herein, we investigated the possibility of culturing cells in capillaries to form the cell layer on its inner wall as chromatographic stationary phase in ACE to screen active compounds.…”

Recent (2018–2020) development in capillary electrophoresis

“…Cells are more suitable than enzymatic proteins to be used in drug screening and characterization by maintaining the natural conformation and bioactivity of proteins. Thus, immobilizing cells to the capillary wall and using them as the drug processor can benefit rapid drug screening and characterization [59]. A method called immobilized cell capillary electrophoresis (ICCE) has been introduced to screen the drugs targeting integrin macrophage antigen-1 (MAC-1, CD11b/CD18) under optimal physiological conditions [60].…”

Screening bioactive compounds from natural product and its preparations using capillary electrophoresis

“…Yu et al. immobilized Chinese hamster ovary cells transfected by overexpressing endothelin receptors on the internal surface of capillary column as the stationary phase for screening and detecting endothelin antagonists from TCMs. Finally, magnolol, geniposide, and honokiol were determined to be the active compounds that have fairly strong antagonism to endothelin.…”