Flow-based combinatorial antibody profiling: an integrated approach to cell characterization

Methods Mol Biol. 2011:699:97-118. doi: 10.1007/978-1-61737-950-5_6.

Abstract

BD FACS™ CAP (CAP = combinatorial antibody profile) is a screening tool for rapid characterization of human cell surface protein expression profiles using semi-automated high-throughput flow cytometry. The current configuration consists of 229 directly conjugated antibodies arrayed in a 96-well plate as three-color cocktails, which enables the characterization of each of the 229 individual surface markers. Each individual cell type of interest is analyzed on the 96-well screening plates and the data are acquired on a flow cytometer equipped with a high-throughput sampler. The expression level of each marker for each cell type is then calculated using semiautomated custom flow cytometry software. The process of characterizing these surface markers in a highly efficient manner using BD FACS™ CAP is enabled by automated liquid handling for staining, automated flow cytometry for data acquisition, and standardized algorithms for automated data analysis.

MeSH terms

  • Antibodies / immunology*
  • Antigens, Surface / genetics
  • Antigens, Surface / immunology
  • Cells, Cultured
  • Cluster Analysis
  • Electronic Data Processing
  • Flow Cytometry*
  • Fluorescent Dyes / metabolism
  • Gene Expression Profiling / instrumentation
  • Gene Expression Profiling / methods*
  • High-Throughput Screening Assays
  • Humans
  • Quality Control
  • Sensitivity and Specificity
  • Software
  • Staining and Labeling

Substances

  • Antibodies
  • Antigens, Surface
  • Fluorescent Dyes