Droplet Split-and-Contact Method for High-Throughput Transmembrane Electrical Recording

Abstract

This paper describes the rapid and repetitive formation of planar lipid bilayers via a mechanical droplet contact method for high-throughput ion channel analysis. In this method, first, an aqueous droplet delivered in a lipid-in-oil solution is mechanically divided into two small droplets. Second, the two small droplets contact each other, resulting in the lipid bilayer formation. Third, an ion channel is immediately reconstituted into the bilayer and the transmembrane current signals are measured. By repeating this procedure, massive data sets of the channel signals can be obtained. This method allowed us to perform statistical analysis of α-hemolysin conductance (n = 256 within 30 min) and channel inhibition experiments by contacting different types of the droplets in a short time frame.

Channel Current Analysis for Pore-forming Properties of an Antimicrobial Peptide, Magainin 1, Using the Droplet Contact Method

“…11,15 Four individual BLMs can be formed simultaneously in this device, which allowed for a higher-throughput measurement compared to the conventional system. Using this device, the DOPC or DOPE/DOPG (3:1 mol/mol) mixture (lipids/n-decane, 10 mg/ mL) solution (2.3 μL) was poured into each chamber.…”

“…11 Using bilayer lipid membranes (BLMs), we have been measuring various membrane protein and biological nanopores, e.g., single molecule detection using DNA aptamers, 14 high-throughput ion channel measurements at the single-protein level, 11,15 solution-exchange experiment in the microdroplet system, 16 and the development of portable nanopore sensors. 17 In this paper, we report on the pore-forming properties of magainin 1 (GIGKFLHSAGKFGKAFVGEIMKS) from channel current conductance measurements using the high-throughput DCM.…”

Hairpin DNA Unzipping Analysis Using a Biological Nanopore Array

“…12 Furthermore, we constructed a droplet array for high-throughput measurements. 13 Using bilayer lipid membranes (BLMs), we measured various membrane proteins and biological nanopore, for example, single molecule detection using DNA aptamers, 14 high-throughput ion channel measurements at the single-protein level, 13,15,16 solutionexchange techniques via the microdroplet system, 17 and the development of portable nanopore sensors. 18 In this study, we Figure 1.…”

Membrane protein-based biosensors

“…Although numbers of the methods and devices have been reported, further development and improvement are required, especially for sensing devices such as usability, stability and portability [149][150][151][152][153][154][155]. Moreover, the integration of sensor elements on a single platform would allow performance of multiplex assays for detection of multiple target analytes and facilitate quantitative analyses [156][157][158][159][160][161].…”