Artificial mycolic acid membranes

Inventors

Gundlach, Jens • Derrington, Ian M. • Langford, Kyle W.

Assignees

WASHINGTON, University of • University of Washington

Abstract

Provided herein are artificial membranes of mycolic acids. The membranes may be unsupported or tethered. These membranes are long lived and highly resistant to electroporation, demonstrating their general strength. The mycolic acid membranes are suitable for controlled studies of the mycobacterial outer membrane and can be used in other experiments, such as nanopore analyte translocation experiments.

Core Innovation

The disclosed invention relates to artificial mycolic acid lipid bilayers spanning an aperture, where the mycolic acid membrane has a rupture voltage greater than 0.5 V and can resist electroporation at an applied 200 mV for more than 24 hours. The document emphasizes that these mycolic acid membranes are long-lived and highly resistant to electroporation, enabling stable bilayers under applied electrical conditions.

The invention further enables formation of stable bilayers that include embedded protein nanopores. The described protein nanopores include MspA and variants, as well as α-hemolysin and OmpATb, enabling functional nanopore behavior within the long-lived mycolic acid bilayers.

The document provides quantitative characterization and functional observations showing that mycolic acid membranes have higher rupture voltages than comparison membranes, and that bilayer lifetime and thickness are consistent with the described membrane behavior. It also describes pH-related reformation and stability behavior, and reports ion-current readouts in which nanopore insertion in the mycolic acid bilayer yields measurable current behavior suitable for analyte translocation readouts.

Claims Coverage

The partial content provided shows three independent claims (clm-00001, clm-00010, clm-00019). Across these, the core inventive coverage is defined by mycolic-acid-based artificial bilayers spanning an aperture with specified electroporation/rupture resistance, optionally including embedded nanopores, and in one case providing a system configuration for analyte translocation and ion-current pattern readouts. Together, these claims cover both the membrane and system, with quantitative electroporation resistance thresholds and optional nanopore embodiments.

The independent claim set is anchored on mycolic-acid artificial bilayers spanning an aperture (or positioned between conductive media) with rupture voltage greater than 0.5 V, and includes an electroporation-resistance limitation at 200 mV for more than 24 hours. Dependent claim information in the provided partial content further indicates nanopore-containing embodiments (including α-hemolysin, Msp, OmpATb, and mutant MspA) and a detection use case involving electrophoretic analyte translocation and measurement of an ion-current characteristic current pattern.

Stated Advantages

Documented Applications