A novel method for patch-clamp automation

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• 作者：D. Vasilyev (1)
  T. Merrill (2)
  A. Iwanow (2)
  J. Dunlop (1)
  M. Bowlby (1)
  
• 关键词：Automation ; Patch ; clamp recordings ; Ion channel ; hERG ; Drug discovery
• 刊名：Pfl眉gers Archiv - European Journal of Physiology
• 出版年：2006
• 出版时间：May 2006
• 年：2006
• 卷：452
• 期：2
• 页码：240-247
• 全文大小：2527KB
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• 作者单位：D. Vasilyev (1)
  T. Merrill (2)
  A. Iwanow (2)
  J. Dunlop (1)
  M. Bowlby (1)
  
  1. Discovery Neuroscience, Wyeth Research, CN 8000, Princeton, NJ, 08543-8000, USA
  2. Biomedical Engineering, Wyeth, CN 8000, Princeton, NJ, 08543-8000, USA
  
• ISSN：1432-2013

文摘

An increasing demand of the pharmaceutical industry for automated electrophysiological stations for ion channel drug discovery has recently resulted in the development of several commercial platforms for secondary and safety screening of ion channel modulators. These commercial systems have demonstrated an enhanced throughput, however, often at the expense of some quality-sensitive aspects of traditional patch-clamp recordings. To improve data quality and content, we have developed a patch-clamp robot that fully automates manual patch-clamp recordings, including patch pipette handling, gigaseal formation, obtaining whole-cell or perforated-cell configuration, drug application, and data acquisition. Utilization of glass micropipettes results in high-quality electrophysiological recordings with an overall success rate of about 30% in perforated-cell mode. A fast drug application system with low volume requirements (1-.5?ml) allows the study of ligand-gated ion channels on a millisecond scale. As proof-of-concept, we present two assays developed for voltage-gated human ether-a-go-go-related and ligand-gated α7 nicotinic receptor ion channels. The system throughput was a single concentration–response curve every 30-0?min or 12-7 6-point concentration–response curves daily, representing a significant improvement of typical manual patch-clamp throughput. This system represents an efficient method for patch-clamp automation without the need for a complex and expensive electrophysiological set-up for cell visualization.