A Cell Microarray Format: A Peptide Release System Using a Photo-Cleavable Linker for Cell Toxicity and Cell Uptake Analysis

Methods Mol Biol. 2016;1352:199-210. doi: 10.1007/978-1-4939-3037-1_15.

Abstract

There has been increasing interest in the potential use of microarray technologies to perform systematic and high-throughput cell-based assays. We are currently focused on developing more practical array formats and detection methods that will enable researchers to conduct more detailed analyses in cell microarray studies. In this chapter, we describe the construction of a novel peptide-array format system for analyzing cellular toxicity and cellular uptake. In this system, a peptide is immobilized at the bottom of a conventional 96-well plate using a photo-cleavable linker. The peptide can then be released from the bottom by irradiating the desired wells with UV light, thus allowing the cytotoxicity or cellular uptake of the peptide to be monitored. This system will facilitate the realization of high-throughput cell arrays for cellomics analyses and cell-based phenotypic drug screens.

Keywords: Cell microarrays; Cellular uptake; Cytotoxicity; Designed peptide; Photo-cleavable linker.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Count
  • Cell Line
  • Cytotoxins / chemistry
  • Cytotoxins / metabolism*
  • Cytotoxins / toxicity*
  • Drug Design
  • Humans
  • Immobilized Proteins / chemistry
  • Immobilized Proteins / metabolism
  • Immobilized Proteins / toxicity
  • Mice
  • Microscopy, Confocal
  • Models, Molecular
  • Peptides / chemistry
  • Peptides / metabolism*
  • Peptides / toxicity*
  • Permeability
  • Protein Structure, Secondary
  • Protein Transport
  • Tissue Array Analysis / methods*
  • Ultraviolet Rays*

Substances

  • Cytotoxins
  • Immobilized Proteins
  • Peptides