Table 2.

Strengths and limitations of currently available multiplexing platforms for biomarker validation

Multiplex PlatformStrengthsWeaknesses
Multiplex Immunoassays
    Suspension arrays
  1. Smaller sample volume and higher throughput than traditional single analyte immunoassays

  2. Many commercially available multiplex assays available from multiple sources

  1. Nonspecific binding of serum proteins directly to microspheres may result in bead aggregation and nonspecific fluorescent emission

  2. Difficult to optimize assay conditions for multiple analytes

    Planar arrays
  1. Smaller sample volume and higher throughput than traditional single analyte immunoassays

  2. Does not require flow cytometric bead analysis

  1. Damage to the capture antibodies by mechanical forces may occur during spotting

  2. Large dynamic range of serum protein abundance limits potential combinations of analyte proteins within an array

  3. Difficult to optimize assay conditions for multiple analytes

Mass spectrometry with selected reaction monitoring
  1. Exquisite specificity

  2. Ability to multiplex hundreds of analytes

  1. Less sensitivity than immunoassays for low abundance proteins

  2. Generation of peptide targets is labor intensive

  3. High abundance proteins may interfere with analyte detection