Ion channel and toxin measurement using a high throughput lipid membrane platform.
Measurements of ion channels are important for scientific, sensing and pharmaceutical applications. Reconstitution of ion channels into lipid vesicles and planar lipid bilayers for measurement at the single molecule level is a laborious and slow process incompatible with the high throughput methods and equipment used for sensing and drug discovery. A recently published method of lipid bilayer formation mechanically combines lipid monolayers self-assembled at the interfaces of aqueous and apolar phases. We have expanded on this method by vertically orienting these phases and using gravity as the driving force to combine the monolayers. As this method only requires fluid dispensation, it is trivially integrated with high throughput automated liquid-handling robotics. In a proof-of-concept demonstration, we created over 2200 lipid bilayers in 3h. We show single molecule measurements of technologically and physiologically relevant ion channels incorporated into lipid bilayers formed with this method.
Poulos
Jason L
JL
Department of Bioengineering, University of California, Los Angeles, USA.
Jeon
Tae-Joon
TJ
Damoiseaux
Robert
R
Gillespie
Eugene J
EJ
Bradley
Kenneth A
KA
Schmidt
Jacob J
JJ
D015374:Biosensing Techniques / Q000295:instrumentation*; D004563:Electrochemistry / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007473:Ion Channels / Q000737:chemistry* / Q000187:drug effects*; D008051:Lipid Bilayers / Q000737:chemistry*; D008567:Membranes, Artificial; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D018675:Toxicity Tests / Q000295:instrumentation* / Q000379:methods; D014118:Toxins, Biological / Q000008:administration & dosage*
D007473:Ion Channels; D008051:Lipid Bilayers; D008567:Membranes, Artificial; D014118:Toxins, Biological