Robotic implementation of assays: tissue-nonspecific alkaline phosphatase (TNAP) case study

Methods Mol Biol. 2013:1053:53-84. doi: 10.1007/978-1-62703-562-0_4.

Abstract

Laboratory automation and robotics have "industrialized" the execution and completion of large-scale, enabling high-capacity and high-throughput (100 K-1 MM/day) screening (HTS) campaigns of large "libraries" of compounds (>200 K-2 MM) to complete in a few days or weeks. Critical to the success these HTS campaigns is the ability of a competent assay development team to convert a validated research-grade laboratory "benchtop" assay suitable for manual or semi-automated operations on a few hundreds of compounds into a robust miniaturized (384- or 1,536-well format), well-engineered, scalable, industrialized assay that can be seamlessly implemented on a fully automated, fully integrated robotic screening platform for cost-effective screening of hundreds of thousands of compounds. Here, we provide a review of the theoretical guiding principles and practical considerations necessary to reduce often complex research biology into a "lean manufacturing" engineering endeavor comprising adaption, automation, and implementation of HTS. Furthermore we provide a detailed example specifically for a cell-free in vitro biochemical, enzymatic phosphatase assay for tissue-nonspecific alkaline phosphatase that illustrates these principles and considerations.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Alkaline Phosphatase / analysis*
  • Automation, Laboratory
  • Biological Assay
  • Cell Line
  • Enzyme Assays / instrumentation*
  • Enzyme Assays / methods*
  • High-Throughput Screening Assays*
  • Humans
  • Robotics / methods*
  • Small Molecule Libraries

Substances

  • Small Molecule Libraries
  • ALPL protein, human
  • Alkaline Phosphatase