Mark E. Sobel, MD, PhD ASIP Executive Officer http://www.asip.org/about/executive_officer.cfm

1. Best Practices of Biobanking and Specimen Collection: Quality Control Systems in Biobanking and Impact on Validity of ResearchResults Lunch and Learn ASIP Annual Meeting at Experimental Biology 2012 April 23, 2012 Mark E. Sobel, MD, PhD ASIP ExecutiveOfficer http://www.asip.org/about/executive_officer.cfm

2. ISBER Best Practices • Biospecimen Science • Quality Control and Proficiency Testing • NCI Best Practices • Human Biological Materials • Special Ethical Considerations Outline

3. Fay Betsou, PhD Integrated Biobank of Luxembourg Presentation at ASIP Annual Meeting at Experimental Biology 2011 Acknowledgments Carolyn Compton, MD Office of Biorepositories and Biospecimen Research National Cancer Institute National Institutes of Health http://biospecimens.cancer.gov/default.asp OBBR slides in thispresentation have been revised

4. International Society for Biological and EnvironmentalRepositories Best Practices for Repositories: Collection, Storage, Retrieval and Distribution of Biological Materials for Research Second Edition: Cell Preservation Technology, Volume 6, Number 1, 2008 Third Edition: Biopreservation and Biobanking Volume 10, Number 2, 2012

5. Biospecimen Science • What is biospecimen science? • Biospecimen Science is the multidisciplinary field of study responsible for establishing tested and proven biospecimen resource-related procedures based on experimentation in the areas of specimen collection, processing, shipping, and storage • Why is it needed? • Biospecimens are composed of active and reactive living cells or cell products, making them highly complex. • The collection, handling, and storage process can profoundly alter the molecular profile and quality of biospecimens. • Such alterations, though artificial, can be misinterpreted as disease related or disease specific. • High degrees of sensitivity and specificity in new molecular techniques raise the bar for analyte (specimen) data and quality.

6. Biospecimen Science • Quality of human biospecimens is multifactorial and is determined by: • Type of specimen: Normal tissue, tumor tissue, serum, plasma • Physical state of the specimen • Amount and type of specimen characterization data • Amount and type of quality control exercised • Amount and type of clinical data • Permitted use of the specimen • The analysis to be performed and the biomolecules targeted by the analysis • The goal of the research (application of the data)

7. Quality Control - definition « the operational techniques and activitiesthat are used to satisfyqualityrequirements », ISO9000:2000 Nestler W. Effect of Quality Control programs on the organizational structure of the hospitals. Bull. NY Acad. Med. 1976;52:157-163 Glenn GC, Hathaway TK. Effects of specimenevaporation on quality control. Am. J. Clin. Pathol. 1976;66:645-652 Calam RR. Reviewing the importance of specimen collection. J. Am. Med. Tech. 1977;39:297-302 « confirmation through provision of objective evidence thatrequirements for a specificintended use or application are fulfilled », ISO9000:2000

8. IntegratedQuality Control Systems QC / Validation Biobank BiobankorResearchLab SampleProcessingMethod Quality Control Assay Method Validation / ProficiencyTesting

9. ISBER ProficiencyTesting Program:Interlaboratorycomparisons to identifyproblems in laboratories and interlaboratorydifferences • Launched in 2011 • DNA Quantification • RNA Integrity and Quantification • Plans for 2012 (to beconfirmed May 2012) • CellViability • Tissue Histology • Tissue Antigenicity

10. in vivoPreanalytical Variables • Position- Fasting- Smoking- Stress- Circadianrhythms- Menstrual cycle- Pregnancy- Physicalexcercise • Caffeine : - phosphodiesterase cAMPdegradation- adrenalinsecretion - triglycleride lipase lipidacidslinked to albumin # of albumin-availablesites

11. in vitroPreanalytical Variables • Collection tubes • shedding of components from the tube • silicones, polymeric surfactants, clotinhibitors or activators, rubberstoppers and plastics • adsorption of serumproteins, RNA, DNA to the tube • Anticoagulants • citrate (dilution) • heparin (binding) • EDTA (plateletclumping) • Clotting time and time of incubation beforecentrifugation • Storage conditions • RT, changes after 8h (m/z<3000) • 4°C, changes after 48h • freeze-thawcycles, controversy (peptide aggregation, precipitation and adsorption)

12. SPREC-01Biospecimenpreanalytical code Standard PreanalyticalCoding for Biospecimens Cancer EpidemiologyBiomarkers and Prevention2010;19:1004-11

13. Ideal QC Biomarkers • Ubiquitous • Measurable by widely accessible methods • On/Off response Stability: capability of a sample material to retain the initial value of a measuredquantityfor a definedperiod of time withinspecificlimitswhenstoredunderdefined conditions ISBER BS WG, Cancer Epidemiol Biomarkers Prevention 2009;18:1017-25

14. Why validate QC methods ? • givefull confidence in samplequality • use samplesafter a long time • provide « referencematerials » • ensurevalidity of researchresults

15. NCI Best Practices for Biospecimen Resources Office of Biorepositories and Biospecimen Research National Cancer Institute National Institutes of Health U.S. Department of Health and Human Services

16. The Premise of Biospecimen Science Quality of human biospecimens is multifactorial and is determined by the: • Type of specimen: Normal tissue, tumor tissue, serum, plasma • Physical state of the specimen • Amount and type of specimen characterization data • Amount and type of quality control exercised • Amount and type of clinical data • Permitted use of the specimen • The analysis to be performed and the biomolecules targeted by the analysis • The goal of the research (application of the data)

17. The Role of Biospecimen Resources in 21st Century Medicine Biospecimen resources encompassing large quantities of high-quality, clinically annotated biospecimens are needed to: • Identify and validate targets for detection, diagnosis, treatment, and prevention • Identify disease mechanisms • Develop a molecular-based taxonomy of cancer • Develop screening tests for biomarkers associated with certain disease subtypes • Group patients for testing of new drugs based on their genetic characteristics and likelihood of positive response • Group patients based on the biomarkers of their disease to determine which treatment is appropriate Biospecimen resources are critical to accelerate the development of molecular-based diagnostics and therapeutics for personalized medicine

18. Key Requirements for Biospecimen Resources inthe Post-Genomic Age • Diversity of cancer types and populations based on continual review of researcher needs • Access through a timely, centralized, peer-review process • Ethical and privacy compliance through a chain of trust • Resources provided without intellectual property restrictions • Pathology and clinical annotation (including longitudinal) • State-of-the-art informatics system to streamline the research production process and create in silicoresearch capabilities • Communication and outreach efforts • Best practice-based and data-driven SOPs to enable reproducible and comparable (additive) results

19. Specimen Collection, Processing, Storage, Retrieval, and Dissemination • Handle specimens as appropriate for specimen type and study design. • Minimize collection/processing time as appropriate. • Develop a comprehensive quality management system to include SOPs to document all protocols and a training program for all appropriate personnel. • Annotate specimens with key collection, processing, and storage data. • Monitor specimen inventory with a tracking system. • Store specimens in a stabilized state without unnecessary thawing/refreezing. Dispose of specimens according to clear rules. • Review and document storage equipment performance on a regular basis. • Follow specimen-appropriate biosafety, packaging, and shipping procedures. • Train personnel to adhere to specimen shipping regulations. Control and monitor required shipping temperature.

20. Collecting/Managing Clinical Data/QA/QC • Collect and store relevant clinical and epidemiologic data associated with a specimen, including longitudinal data, if applicable. • Use an informatics system that tracks all aspects of collection, processing, and distribution. • Comply with applicable privacy rules and human subjects regulations. Collecting and Managing Clinical Data Quality Assurance/Quality Control • Adhere to a written quality management system that describes QA and QC procedures. • Maintain QA/QC training records for personnel. • Adhere to and periodically review SOPs. • Have security systems in place, including alarms and backup power. • Include a computerized inventory tracking system in the data management plan. • Develop a facility disaster plan. • Maintain all equipment properly according to SOPs.

21. Biosafety • Assume that all specimens are potentially infectious – provide appropriate vaccines, e.g., hepatitis. • Adhere to governmental and accrediting agency requirements. • Identify and address biosafety risks. • Record exposure incidents and provide personnel with appropriate treatment. • Establish indemnification agreements with users of biospecimens (except where prohibited by law). • Develop additional policies and procedures as appropriate for chemical, electrical, fire, occupational, and radiological safety. Biosafety

22. Biospecimen Resource Informatics • Assign a unique identifier (number and/or barcode) to each specimen. • Update the biospecimen resource database each time the specimen is moved or modified. • Use informatics systems that support the linking of specimens with associated data and protect the health information of patients. • Adhere to or initiate review of NCI Center for Bioinformatics guidelines and tools; caBIG™ “silver-level” compatibility is recommended. Biospecimen Resource Informatics: Data Management, Inventory Control, and Tracking

23. Access to Biospecimens and Data • Develop clear policies for specimen and data access. • Develop clear guidelines for sample distribution and clinical data sharing (note: NCI Best Practices state that protocol-specific requirements should be met before other access is considered). • Ensure that investigators have timely, equitable, and appropriate access, without undue administrative burden. • Charge for samples only to recover costs. • If a biospecimen resource needs to close, announce the availability of specimens for transfer. • Restrict access to subjects’ identities and medical, genetic, social, and personal histories via data access system with defined privilege levels.

24. Privacy Protection/Custodianship • Protect the privacy of information and follow applicable regulations. • Follow documented policies on employee access to data or specimens. • Provide levels of security that are appropriate to the type of biospecimen resource. Privacy Protection Custodianship • Include plans for custodianship of collected specimens and associated data in biospecimen resource protocols. • Develop plans to handle/dispose of specimens and associated data: • At end of the budget period of the grant • At completion of the specific research objectives of the study • Identify and disclose financial conflicts of interest. • In informed consent language, disclose that specimens may help to develop products, tests, or discoveries that may have commercial value.

25. Types of Human Biological Materials • Germline vs. Somatic cell • Germline • Inheritability • Implications for immediate and extended family • Implications for ethnic group • Use of “normal” tissues • Somatic cell • Acquired mutations • Use of diseased tissues • No implications for family

26. Types of Human Biological Materials • Unidentifiable vs Identifiable • Unidentifiable • Anonymous • Anonymized • Identifiable • Coded (Linked) • Identified

27. Ethical PrinciplesThe Belmont Report • Respect for persons (personal autonomy) • Beneficence • Justice

28. Federal Regulations • Department of Health and Human Services Common Rule, 45 CFR 46 • Food and Drug Administration: 50 CFR 56 • Institutional Review Boards • Informed consent • OHRP: Office of Human Research Protections • HIPAA- April, 2003 • Inconsistencies between HIPAA and the Common Rule

29. Human Subjects Protection:Use of Human Biological Materials • HBMs include: • Tissue samples • Blood, sputum, urine, bone marrow, etc. • Freshly obtained and archived materials • HBMs are subject to the same regulations as human subjects directly enrolled in studies • Therefore, informed consent and approval by an IRB may be required before using HBMs

30. Identifiable HBMs- The Common Rule • Any HBM that can be identified by any one person, anywhere, is an identifiable sample • If a sample is coded, and any investigator keeps a key to the code, the sample is identifiable • Exception: If the recipient of the HBMs signs an agreement that there is no intent to identify the samples, the sample may be considered unidentifiable.

31. Definition of a Human Subject-The Common Rule • Does NOT include: • Deceased persons (autopsy specimens) • Publicly available information • Unidentifiable (Anonymous, Anonymized) Samples

32. Waivers of Informed Consent-Common Rule An IRB may grant a waiver of informed consent under the Common Rule (not the FDA) if four criteria are met: • Minimal risk • Respect for autonomy and the rights of the individual • Impracticable • Notification

33. Waivers of Informed Consent-Common Rule • Waivers are rarely granted for identified samples • Waivers are usually (but not always) granted for collecting anonymous samples • Waivers are usually (but not always) granted for using anonymous or anonymized samples • Waivers are occasionally granted for coded (linked) samples: • Germline • Somatic cell

34. OHRP Guidance: Components of Repositories Repository activities involve three components, each of which must satisfy certain requirements: • collectors of tissue samples • repository storage and data management center • recipient investigators

35. OHRP Repository Guidance Tissue collector Recipient Investigator Repository Storage and Data Management Center Tissue collector Recipient Investigator Tissue collector Recipient Investigator IRB Review Informed Consent Submittal Agreement Assurance of Compliance IRB Review SampleInformedConsent Certificate of Confidentiality Assurance of Compliance RecipientAgreement Local Policies

36. Ethical Principles for the 21st Century • Respect for persons (personal autonomy): • Informed consent • Respect for privacy and confidentiality • Beneficence • Justice