Leveraging Automation toward Development of a High-Throughput Gene Expression Profiling Platform

SLAS Discov. 2021 Mar;26(3):410-419. doi: 10.1177/2472555220956593. Epub 2020 Sep 16.

Abstract

We previously developed a panel of one-step real-time quantitative reverse transcription PCR (one-step qRT-PCR; hereafter referred to as qRT-PCR) assays to assess compound efficacy. However, these high-cost, conventional qRT-PCR manual assays are not amenable to high-throughput screen (HTS) analysis in a time-sensitive and complex drug discovery process. Here, we report the establishment of an automated gene expression platform using in-house lysis conditions that allows the study of various cell lines, including primary T cells. This process innovation provides the opportunity to perform genotypic profiling in both immunology and oncology therapeutic areas with quantitative studies as part of routine drug discovery program support. This newly instituted platform also enables a panel screening strategy to efficiently connect HTS, lead identification, and lead optimization in parallel.

Keywords: automation; biomarkers; drug discovery; gene expression; one-step qRT-PCR.

Publication types

  • Validation Study

MeSH terms

  • Antigens, Neoplasm / genetics
  • Antigens, Neoplasm / immunology
  • Automation, Laboratory / instrumentation
  • Automation, Laboratory / standards*
  • Baculoviral IAP Repeat-Containing 3 Protein / genetics
  • Baculoviral IAP Repeat-Containing 3 Protein / immunology
  • Cell Line, Tumor
  • Chemokine CCL3 / genetics
  • Chemokine CCL3 / immunology
  • Drug Discovery / methods
  • Gene Expression Profiling / instrumentation
  • Gene Expression Profiling / methods
  • Gene Expression Profiling / standards*
  • Gene Expression Regulation
  • HCT116 Cells
  • High-Throughput Screening Assays / instrumentation
  • High-Throughput Screening Assays / methods*
  • Humans
  • Membrane Proteins / genetics
  • Membrane Proteins / immunology
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / immunology
  • Osteoblasts / cytology
  • Osteoblasts / metabolism
  • Primary Cell Culture
  • Real-Time Polymerase Chain Reaction / methods*
  • Real-Time Polymerase Chain Reaction / standards
  • Ribosomal Proteins / genetics
  • Ribosomal Proteins / immunology
  • T-Lymphocytes / cytology
  • T-Lymphocytes / metabolism

Substances

  • Antigens, Neoplasm
  • CCL3 protein, human
  • CTAG1B protein, human
  • Chemokine CCL3
  • MAGEA4 protein, human
  • Membrane Proteins
  • Neoplasm Proteins
  • Ribosomal Proteins
  • ribosomal protein L32
  • BIRC3 protein, human
  • Baculoviral IAP Repeat-Containing 3 Protein