1. Patient Biospecimen Preanalytics:
Even the Perfect Test Can Give the Wrong Result If the Analyte Is Compromised
Carolyn Compton, MD, PhD
Professor Life Sciences, Arizona State University
Professor Laboratory Medicine and Pathology, Mayo Clinic
Adjunct Professor of Pathology, Johns Hopkins
CMO, National Biomarker Development Alliance
CMO, Complex Adaptive Systems Institute
Scottsdale, AZ
National Academies of Sciences, Engineering, and Medicine
Return of Research Results Workshop
Washington, DC
September 6, 2017

2. Bias and Biospecimens
Sources of Bias in Molecular Marker Research in Cancer
- David F. Ransohoff and Margaret L. Gourlay, 2010

3. Translational Research
Biospecimen Quality Affects Both Molecular Medicine and Translational Research
Analysis Data
Decision Making
DETERMINES QUALITY HERE
Clinical Medicine or
Translational Research
QUALITY HERE
Biospecimen Analysis
Biospecimen Collection
Biospecimen processing and ba
Preanalytic Biospecimen Handling and Processing

4. Pre-analytical Factors Affect Both Molecular Composition and Molecular Quality
Specimen is viable
and biologically reactive
Molecular composition subject to further alteration/degradation
Factors (examples):
Antibiotics
Other drugs
Type of anesthesia
Duration of anesthesia
Arterial clamp time
Factors (examples):
Time at room temperature
Temperature of room
Type of fixative
Time in fixative
Rate of freezing
Size of aliquots
Time 0
ANALYS I S
STORAGE
Patient
Pre-Op
Acquisition
Handling/
Processing
Storage
Distribution
Scientific
Analysis
Medical/
Surgical
Procedures
Restocking
Unused
Sample
Pre-acquisition
Post-acquisition

5. pMAPK IHC of Colon Cancer : HER2 IHC and FISH in Breast Cancer:
Does It Matter?
Cold Ischemia Time and Molecular Assay Results
pMAPK IHC of Colon Cancer :
HER2 IHC and FISH in Breast Cancer:
Loss of Biomarker Signal
Gain of Biomarker Signal
30 min
10 min
HER2/CEP17 = 0.98 a
20 min
4 hr delay
2 hr
60 min c
Khoury T, et al., Mod Pathol Nov;22(11):
Hartmut Juhl, Indivumed GmbH, BRN

6. Blood Collection and Plasma Processing: Biomarkers and Circulating Tumor Cells
Procedure,
Temperature
and Time
Blood Draw
Procedure
Collection
Tubes and
Order of
draw
Distribution
& Storage
Molecular
Analysis
Patient
Consent
and
Preparation
Blood draw: peripheral vein vs. central line
Specimen type: plasma vs. serum
Tube type
Tube fill level
Tube inversions
Time to processing
Centrifuge speed
Temperature variation before and during processing
Storage conditions and freeze-thaw cycles
Transport conditions

7. Analysis in the Clinical Lab: How Is It Assured?
Place where test is done
CLIA/CAP laboratory accreditation
People doing the test
Education
Proficiency testing
Licensure
Platforms used for testing
CDRH approved devices
Processes followed for testing
SOPs
Quality management
Patient samples to be tested
Variably variable: uncontrolled
More is known about the quality of beef in the supermarket
than is known about the quality of human biospecimens in research

8. Impact of Biospecimen Quality on Analysis Data Quality
Effects on Clinical Care
Potential for incorrect diagnosis
Potential for incorrect treatment
Therapy linked to diagnostic test on a biospecimen
Effects on Research
Irreproducible results
Variation in mutation data
Variation in gene expression data
Misinterpretation of artifacts as biomarkers
Return of bad data to patients?

9. The National Academies of Sciences, Engineering and Medicine (IOM) 2016
Biomarker Tests for Molecularly Targeted Therapies:
Key to Unlocking Precision Medicine 
National Academies Press, Jun 30, 2016 
One of the 10 stated goals is to specifically address biospecimen quality.
Goal 9: Enhance specimen handling and documentation to ensure patient safety and the accuracy of biomarker test results. 
“The reliability of biomarker test results depends on the quality of the patient specimens.”
“Professional organizations and health care institutions should develop and implement standards for obtaining adequate specimens.”

10. Powerful Tools: Powerful Risks
Analysis technologies are powerful than ever.
The technological capacity exists to produce low-quality data from low- quality analytes with unprecedented efficiency.
It is now possible to get the wrong answer with unprecedented speed.
Even the best technology cannot spin straw into gold – you must begin with gold.
If garbage has gone in, results must not go out - to either patients or physicians

11. Patient Biospecimen Preanalytics:
Even the Perfect Test Can Give the Wrong Result If the Analyte Is Compromised
Carolyn Compton, MD, PhD
Professor Life Sciences, Arizona State University
Professor Laboratory Medicine and Pathology, Mayo Clinic
Adjunct Professor of Pathology, Johns Hopkins
CMO, National Biomarker Development Alliance
CMO, Complex Adaptive Systems Institute
Scottsdale, AZ
National Academies of Sciences, Engineering, and Medicine
Return of Research Results Workshop
Washington, DC
September 6, 2017