Translating pharmacodynamic biomarkers from bench to bedside: analytical validation and fit-for-purpose studies to qualify multiplex immunofluorescent assays for use on clinical core biopsy specimens.
Multiplex pharmacodynamic (PD) assays have the potential to increase sensitivity of biomarker-based reporting for new targeted agents, as well as revealing significantly more information about target and pathway activation than single-biomarker PD assays. Stringent methodology is required to ensure reliable and reproducible results. Common to all PD assays is the importance of reagent validation, assay and instrument calibration, and the determination of suitable response calibrators; however, multiplex assays, particularly those performed on paraffin specimens from tissue blocks, bring format-specific challenges adding a layer of complexity to assay development. We discuss existing multiplex approaches and the development of a multiplex immunofluorescence assay measuring DNA damage and DNA repair enzymes in response to anti-cancer therapeutics and describe how our novel method addresses known issues.
Pharmacodynamics Assay Development and Implementation Section, Laboratory of Human Toxicology and Pharmacology, Applied/Developmental Research Directorate, Leidos Biomedical Research, Inc, Frederick National Laboratory for Cancer Research, Frederick, MD.
Allison
A
Marrero
Pharmacodynamics Assay Development and Implementation Section, Laboratory of Human Toxicology and Pharmacology, Applied/Developmental Research Directorate, Leidos Biomedical Research, Inc, Frederick National Laboratory for Cancer Research, Frederick, MD.
Scott
S
Lawrence
Pharmacodynamics Assay Development and Implementation Section, Laboratory of Human Toxicology and Pharmacology, Applied/Developmental Research Directorate, Leidos Biomedical Research, Inc, Frederick National Laboratory for Cancer Research, Frederick, MD.
Deborah
D
Wilsker
Pharmacodynamics Assay Development and Implementation Section, Laboratory of Human Toxicology and Pharmacology, Applied/Developmental Research Directorate, Leidos Biomedical Research, Inc, Frederick National Laboratory for Cancer Research, Frederick, MD.
Andrea Regier
AR
Voth
Pharmacodynamics Assay Development and Implementation Section, Laboratory of Human Toxicology and Pharmacology, Applied/Developmental Research Directorate, Leidos Biomedical Research, Inc, Frederick National Laboratory for Cancer Research, Frederick, MD. Electronic address: kindersr@mail.nih.gov.
Robert J
RJ
Kinders
D000970:Antineoplastic Agents / Q000493:pharmacokinetics* / Q000627:therapeutic use; D014408:Biomarkers, Tumor / Q000032:analysis*; D062005:Biopsy, Large-Core Needle; D002138:Calibration; D002624:Chemistry, Clinical / Q000379:methods; D004260:DNA Repair / Q000187:drug effects / Q000502:physiology; D004798:Enzymes / Q000032:analysis / Q000378:metabolism; D005455:Fluorescent Antibody Technique / Q000379:methods* / Q000592:standards; D006801:Humans; D007091:Image Processing, Computer-Assisted; D009369:Neoplasms / Q000188:drug therapy / Q000473:pathology*; D040081:Protein Array Analysis / Q000379:methods; D011786:Quality Control; D015203:Reproducibility of Results
D000970:Antineoplastic Agents; D014408:Biomarkers, Tumor; D004798:Enzymes
Assay validation; Cancer drug; Clinical samples; Fitness-for-purpose; Multiplex assay; Targeted therapy
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