Assay Development

Development and validation of multiplexed immunoassays is a core competency of Rules-Based Medicine, a Q2 Solutions Company. We first select the commercially available antigens and antibodies for each target analyte. If none are found, the necessary antigen and antibody development is outsourced to one of several reliable vendors. All Multi-Analyte Profiles (MAPs) are developed and validated using a standard procedure based upon the simple principles of immunoassays. Each individual assay is first developed as a single-plex test to achieve the sensitivity and dynamic range necessary to quantify the target analyte from the desired sample type. The resulting single-plex assays are then combined into a multiplex. Multiplexed calibrators (8 levels per analyte) and controls (3 levels per analyte) are developed. The multiplexed assays, calibrators, and controls are validated with a battery of analytical procedures to complete the development process. Validation parameters that are determined for each multiplexed analyte assay include:

  • Least detectable dose (LDD)
  • Normal range
  • Dynamic range
  • Imprecision
  • Spiked recovery
  • Parallelism
  • Correlation (where possible)
  • Cross-reactivities
  • Matrix interferences

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