UV-DIB: label-free permeability determination using droplet interface bilayers. Issue 5 (2nd February 2022)

• Record Type: Journal Article
• Authors: Strutt, Robert; Sheffield, Felix; Barlow, Nathan E.; Flemming, Anthony J.; Harling, John D.; Law, Robert V.; Brooks, Nicholas J.; Barter, Laura M. C.; Ces, Oscar
• Is Part Of: Lab on a chip. Volume 22:Issue 5(2022)
• Page Start: 972
• Page End: 985
• Publication Date: 2022-02-02
• Subjects: Miniature electronic equipment -- Periodicals; Combinatorial chemistry -- Periodicals; Biotechnology -- Periodicals; 543.0813
• DOI: 10.1039/d1lc01155c
• Languages: English
• ISSNs: 1473-0197

Abstract

Abstract : Label-free permeability measurement in droplet interface bilayers to assess the relationship between bilayer lipid composition and permeant chemical structure. Abstract : Simple diffusion of molecular entities through a phospholipid bilayer, is a phenomenon of great importance to the pharmaceutical and agricultural industries. Current model lipid systems to probe this typically only employ fluorescence as a readout, thus limiting the range of assessable chemical matter that can be studied. We report a new technology platform, the UV-DIB, which facilitates label free measurement of small molecule translocation rates. This is based upon the coupling of droplet interface bilayer technology with implemented fiber optics to facilitate analysis via ultraviolet spectroscopy, in custom designed PMMA wells. To improve on current DIB technology, the platform was designed to be reusable, with a high sampling rate and a limit of UV detection in the low μM regime. We demonstrate the use of our system to quantify passive diffusion in a reproducible and rapid manner where the system was validated by investigating multiple permeants of varying physicochemical properties across a range of lipid interfaces, each demonstrating differing kinetics. Our system permits the interrogation of structural dependence on the permeation rate of a given compound. We present this ability from two structural perspectives, that of the membrane, and the permeant. We observed a reduction in permeability between pure DOPC and DPhPC interfaces, concurring with literature and demonstrating our ability to study the effects of lipid composition on permeability. In relation to the effects of permeant structure, our device facilitated the rank ordering of various compounds from the xanthine class of compounds, where the structure of each permeant differed by a single group alteration. We found that DIBs were stable up to 5% DMSO, a molecule often used to aid solubilisation of pharmaceutical and agrochemical compounds. The ability of our device to rank-order compounds with such minor structural differences provides a level of precision that is rarely seen in current, industrially applied technologies.