What have we learned about Biomarkers

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What have we learned about Biomarkers?

• Paul Rhyne
• Bristol-Myers Squibb Company

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Outline

• Biomarker classifications
• Validation considerations
• Performing Biomarker Assays
• Biomarker assay format
• Summary

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Biomarker Classifications

• Exploratory Biomarkers
• Used to generate hypotheses
• Minimal Validation
• Diagnostic prognostic Biomarkers
• Disease outcome and efficacy
• Advanced Validation
• Safety
• Access toxicity to organs/systems
• Biomarker data used to assess eligibility,
  treatment, discontinuation etc.
• Advanced Validation
• Decision making
• Established biomarkers
• Determine how well a drug works on the target
• Advanced Validation
• Surrogate
• Substitute for clinical endpoints in drug
  efficacy studies
• Full Validation

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Outline

• Biomarker classifications
• Validation considerations
• Performing Biomarker Assays
• Biomarker assay format
• Summary

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Biomarker Assay Validation

• Sensitivity and assay reproducibility
• What level is needed?
• Bias
• Difference between measured and the true value
• Sample type and handling
• Interference/matrix
• Biomarker stability
• Specificity
• Assay
• Clinical Ex. How well the biomarker assay
  correctly identifies diseased patients from
  healthy
• Assay stability and reproducibility (Lot to lot)
• Stability of technology manufacturer

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Outline

• Biomarker classifications
• Validation considerations
• Performing Biomarker Assays
• Biomarker assay format
• Summary

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Performing Biomarker assays(In house vs
Outsource)

• Equipment and Instrumentation
• Biomarker Assay expertise
• Data handling
• Location of clinical site vs analysis site
• Technology support for assay
• Commercial assay vs in-house assay

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Outline

• Biomarker classifications
• Validation considerations
• Performing Biomarker Assays
• Biomarker assay format
• Summary

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Immunoassays

• Solid phase based immunoassays
• Microtiter plate based (ELISA)
• Spotted arrays
• Bio-chips
• Suspension arrays
• ECL based detection

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Microtiter plate based (Ex. ELISA)
Biotinylated reporter antibody
Sample
Streptavidin HRP
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Spotted arrays (Ex. Searchlight)
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Suspension array (Ex. Luminex)
Dye 2
Dye 1
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Bead based Immunoassay
Streptavidin PE
Biotin 2nd antibody
Captured Biomarker
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ECL based detection (Ex. MSD)

• The electrochemical reaction occurs within the
  plate and light is measured through a CCD camera
  or photodiodes.
• Working and counter electrodes separated by
  dielectric
• Dielectric also forms physical and surface
  tension barrier

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Lower Limit of Quantitation (LLQ)

• The lowest concentration in the assay for which
  the coefficient of variation is less than 30.
• The lowest concentration in the assay for which
  the measured value is within 30 of true value
  (accuracy).

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Singleplex
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Multiplex
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Outline

• Biomarker classifications
• Validation considerations
• Performing Biomarker Assays
• Biomarker assay format
• Summary

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Summary

• Many types of Biomarkers
• Validation should be fit for purpose
• Biomarker work In house vs outsource?
• Traditional vs newer technology for Biomarker
  Assays

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Acknowledgements

• Giovanni Rizzi
• Yan J. Zhang
• Holly Palme
• Joan Kelsey
• Flora Berisha
• Fizal Nabbie
• Allen Wong
• Russell Weiner

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Thanks!
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Backup slides
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Luminex Multiplex vs Singleplex
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Analysis of Plasma
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Comparative analysis of IP-10
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Comparative analysis of IL-8
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Analysis of IL-1b plasma
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Analysis of TNFa in plasma