title: Robotic implementation of assays: tissue-nonspecific alkaline phosphatase (TNAP) case study.
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.
authors.affiliation: Conrad Prebys Center for Chemical Genomics, Sanford-Burnham Medical Research Institute, La Jolla, CA, USA.
authors.forename: Thomas D Y
authors.initials: TD
authors.lastname: Chung
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis*; D057205:Automation, Laboratory; D001681:Biological Assay; D002460:Cell Line; D057075:Enzyme Assays / Q000295:instrumentation* / Q000379:methods*; D057166:High-Throughput Screening Assays; D006801:Humans; D012371:Robotics / Q000379:methods*; D054852:Small Molecule Libraries
chemical_list: D054852:Small Molecule Libraries; C548267:ALPL protein, human; D000469:Alkaline Phosphatase