Electrophysiological measurement of ion channels is difficult because the ion channel needs to reside in a lipid bilayer membrane for determination of its conductance. The PI's group has developed an artificial lipid bilayer membrane formation technology that enables inexpensive cell-free measurement of ion channels on inexpensive low-tech plastic consumable plates. The proposed innovation is able to measure ion channels from any cell type or source including primary cells and subcellular organelles. Ion channel measurements have already been validated with measurements of conductance modulation of physiologically relevant ion channels by known active pharmaceutical compounds. The proposed work concerns development of a 48 well array plate for this measurement compatible with solution exchange, which would allow rapid parallel screening of ion channels against potential pharmaceutical interactions, and the demonstration of the parallel processing and measurement of membrane arrays by automated instrumentation. Prospective end users of the technology will be queried regarding their willingness to try ion channel screening in a cell-free system, what features would be necessary for that to happen, and attractiveness of unique features of the proposed technology.
Ion channels play key roles in cardiac and neural activity and their disorders have been implicated in epilepsy and cystic fibrosis and a number of other diseases. Because of their central importance, ion channels are critical drug targets - 15 of the top 100 selling drugs are ion channel modulators with a combined market value of $15 billion. Automated Patch Clamp (APC) is the state of the art technology for electrophysiological screening of ion channel activity in cells for drug discovery and safety screening. However, it is well recognized to suffer from low throughput, expensive consumables, and compatibility with limited cell types, and these shortcomings limit the number and scope of drug discovery studies. The proposed cellfree technology has the potential to address all of these shortcomings, thus allowing drugs to be discovered and produced more efficiently and for lower cost, enabling more and better drugs to be produced. The ease of use and low cost of the proposed technology also has the potential to positively affect scientific research and smaller scale laboratory studies of ion channels.
Intellectual Merit: Electrophysiological measurement of ion channels is difficult because the ion channel needs to reside in a lipid bilayer membrane for determination of its conductance. We have developed an artificial lipid bilayer membrane formation technology that enables inexpensive cell-free measurement of ion channels on inexpensive low-tech plastic consumable plates. We constructed an 8 element, automated array aparatus for simultaneous formation of artificial membranes and meausurement of ion channels in them. This proof-of-concept device shows the potential for this technology to allow rapid parallel screening of ion channels against potential pharmaceutical interactions with automated instrumentation. Prospective end users of the technology were queried regarding their willingness to try ion channel screening in a cell-free system, what features would be necessary for that to happen, and attractiveness of unique features of the proposed technology. We found considerable interest in the system, provided that 32-48 wells were measureable in parallel. Broader Impact: Ion channels play key roles in cardiac and neural activity and their disorders have been implicated in epilepsy and cystic fibrosis and a number of other diseases. Because of their central importance, ion channels are critical drug targets—15 of the top 100 selling drugs are ion channel modulators with a combined market value of $15 billion. Automated Patch Clamp (APC) is the state of the art technology for electrophysiological screening of ion channel activity in cells for drug discovery and safety screening. However, it is well recognized to suffer from low throughput, expensive consumables, and compatibility with limited cell types, and these shortcomings limit the number and scope of drug discovery studies. Our cell-free technology has the potential to address all of these shortcomings, thus allowing drugs to be discovered and produced more efficiently and for lower cost, enabling more and better drugs to be produced. The ease of use and low cost of the proposed technology also has the potential to positively affect scientific research and smaller scale laboratory studies of ion channels.
