6/5/2014 Pre‐Analytical Quality Management of Biospecimen: Assurance is good. Control is better. Assurance is good. Control is better. “Inacceptable sample quality due to failures in the pre‐analytical phase and/or improper sample storage in retrospective research studies is a widely known challenge in biomedical R&D.” Prof. Dr. Kurt Zatloukal, Coordinator of a pan‐European Research Infrastructure on Biobanking and Biomolecular Resources (BBMRI) BLOOD WITHDRAWAL ‣ ‣ ‣ ‣ ‣ ‣ ‣ ‣ ‣ Product Launch Booth #112 May 22th, 2014 Beate Kamlage, metanomics GmbH Bianca Bethan, metanomics GmbH BLOOD PROCESSING Sample type Agitation Microclotting Tube underfilled Wrong sequence in sampling Mislabelled Blood draw systems Hemoconcentration due to tourniquet Patient‐related confounders (e. g. fasting, age, gender, BMI, diseases, medication) PLASMA PROCESSING ‣ Time from blood draw to centrifugation* ‣ Temperature from blood draw to centrifugation* ‣ Hemolysis* ‣ Agitation ‣ Centrifugation force ‣ Centrifugation temperature ‣ Remaining blood cells in plasma ‣ Plasma processing/storage time* ‣ Plasma processing/storage temperature* ‣ Sample type errors ‣ Freeze‐thaw‐cycles Oliver Schmitz, metanomics GmbH Philipp Schatz, Metanomics Health GmbH Red: covered by MxP® Quality Control Plasma * Reported to have highest impact on the plasma metabolome (Kamlage et al. 2014 Clin Chem 60, 399‐412) Metanomics Health – a BASF Group Company 2 Assurance is good. Control is better. An Example from Real Life Special Studies: Examples of Metabolite Response to Pre‐Analytical Variations 2 Color by batch of samples 1 2 3 4 5 6 7 8 9 10 11 12 13 14 QUALITY MARKER PATTERN intra‐sample ratio of 2 metabolites 4 3.5 3 2.5 2 1.5 1 *** Serotonin, log10 ng/mL 5 4.5 *** *** 1.5 1 0.5 0 ‐0.5 ‐1 ‐1.5 Shape by group 0.5 Disease Y Control 0 50 100 150 200 250 300 350 400 SAMPLE STORAGE TIME (weeks) Sphingosine‐1‐phosphate (d18:1), log10 ratio versus MxPool™ Samples for a major biomarker study were received from a biobank Identification of a batch of samples (colored in red) which was exposed to adverse pre‐analytical conditions Metanomics Health – a BASF Group Company Color by Confounder *** Control Prolonged blood processing at room temperature ‐0.1 ‐0.2 Prolonged blood processing on ice *** *** Shear force induced hemolysis ‐0.3 ANOVA versus control group, FDR < 0.2 ‐0.4 *** P‐value < 0.0001 ‐0.5 ‣ Serotonin and sphingosine‐1‐phosphate levels in plasma are influenced by pre‐analytical blood processing ‣ Effects can be biochemically interpreted by platelet and erythrocyte metabolism 3 Metanomics Health – a BASF Group Company 4 Metanomics Health – a BASF Group Company 1 6/5/2014 P® Quality Control Plasma MxP® Quality Control Plasma Results from Beta‐Test Result 1: MxP QC Collection Tube Control (data from Bayer Pharma AG) Metabolite levels in plasma allow very comprehensive and sensitive read‐out of human plasma sample quality (quality control) MxP Quality Control Plasma controls EDTA plasma quality at three checkpoints RESULT 1 MxP QC ection Tube Control 1 RESULT 3 MxP QC Blood Processing Control MxP QC Plasma Processing Control 100 10 MxP Quality Score (QS) RESULT 2 Y 1000 0.1 Differences in time and temperature OOD WITHDRAWAL Pipettes / skill of user 0.001 BLOOD PROCESSING PLASMA PROCESSING SAMPLE STORAGE Metanomics Health – a BASF Group Company Negative 18 2 Positive 2 98 Specificity: 90 % Sensitivity: 98 % 1 10 Separation of different sample types by three compounds: citrate plasma, EDTA plasma, h heparin plasma, and serum. Blinded study by Bayer Pharma AG with human blood samples addressing the following confounders in addition to a control group: ‣ Blood storage at room temperature for 6 h ‣ Plasma storage at room temperature for 6, 24, 48 h ‣ Five freeze‐thaw‐cycles of plasma samples Metanomics Health – a BASF Group Company P® Quality Control Plasma Results from Beta‐Test MxP® Quality Control Plasma Report lt 2: MxP QC d Processing Control MxP Collection Tube Control: Result 3: MxP QC Plasma Processing Control 28 0 26 8 6 4 AUC = 0.91 p < 0.001* 2 0 8 AUC = 0.62 p < 0.05* 6 AUC = 1.0 p < 0.001* 4 2 Quality control metabolite concentration in plasma [µmol/L] 2 Checks for EDTA plasma versus citrate/heparin plasma or serum MxP Blood Processing Control: High quality: centrifugation within 2 h Medium quality: centrifugation within 2 to 6 h Low quality: centrifugation > 6 h or platelet activation 24 22 20 18 16 AUC = 0.83 p < 0.001* 14 MxP Plasma Processing Control: 12 High quality: processing time at room temperature within 6 h Medium quality: processing time at room temperature within 6 to 24 h Low quality: processing time at room temperature > 24 h 10 8 6 AUC = 1.0 p < 0.001* 4 2 0 15 min / 21 °C (control) 2 h 0 °C 2 h 21 °C 6 h 0 °C Time of blood incubation / temperature 0 15 min / 21 °C (control) 6 h 21 °C 24 h 21 °C Time of plasma incubation / temperature ots of metabolic quality markers in plasma increasing when EDTA blood (left) or plasma (right) is ated for prolonged time. AUC = Area under curve of ROC versus control. * P‐value of linear mixed or simple (right) model relative to control. Metanomics Health – a BASF Group Company Values are combined into the MxP Quality Score 8 Confounders (100) X 1000 6 Control (20) 0.001 100 Storage time and temperature MxP QC test result 0.1 Z Types of additives in the blood collection tubes Sensitivity and specificity of MxP QC Results 2 and 3 (data from Bayer Pharma AG) Metanomics Health – a BASF Group Company 6/5/2014 MxP® Quality Control Plasma Exclusive ISBER Member Discount P® Quality Control Plasma Offer A) SERVICE MODEL B) LICENSE MODEL Attractive pricing aligned with your quality management budget Cost‐effective solution for upgrading your clinical trial/biobanking quality management; economic for > 500 samples with existing GC‐MS device Send the samples on dry ice to Metanomics Health Analysis is done by Metanomics alth or their affiliates/subcontractors Get the report and result list indicating the quality status of your samples DQUARTERS U.S. OFFICES JAPAN OFFICES nomics Health GmbH er Weg 33 9 Berlin many BASF Plant Science LP 26 Davis Drive Durham, NC 27709 U.S.A. BASF Japan Ltd. Roppongi Hills Mori Tower 21F6‐10‐1 Roppongi, Minato‐ku Tokyo 106‐6121 Japan Metanomics Health – a BASF Group Company Deadline Discount 30 % August 31, 2015 10 % August 31, 2015 Eligible for all out‐licensing fees including IP and sample licenses 10 % MxP® QC Plasma as cost‐effective sample quality check for single projects 10 cipate in our raffle! e your card at Metanomics Health booth (#112) and participate in drawing of a test run of MxP® Quality Control Plasma service equivalent to amples. Discount September 30, 2014 Eligible for all fee‐for‐service projects of ISBER members Create your own report using software provided by Metanomics Health nk you B) LICENSE MODEL Deadline Eligible for pilot‐projects of ISBER members Conduct the analysis at your own facility with your GC‐MS experienced staff Metanomics Health – a BASF Group Company e: +49 30 34807 400 @metanomics‐health.de .metanomics‐health.de A) SERVICE MODEL Metanomics Health – a BASF Group Company MxP® QC Plasma as part of routine protocol for quality assurance in clinical trials erences Vaught J et al. Journal of the National Cancer Institute Monographs 2011;2011:1‐7. Kamlage B et al. Clin Chem 2014;60:399‐412. Tuck MK et al. J Proteome Res 2009;8:113‐7. Yin P et al. Clin Chem 2013;59:833‐45. Yang W et al. Anal Chem 2013;85:2606‐10. Oddoze C et al. Clin Biochem 2012;45:464‐9. Rai AJ and Vitzthum F. Expert Review of Proteomics 2006;3:409‐26. Aguilar‐Mahecha A et al. PLoS One 2012;7:e38290. Greystoke A et al. Ann Oncol 2008;19:990‐5. Hebels DGAJ et al. Environ Health Perspect 2013;121:480‐7. Becker N and Lockwood CM. Clin Biochem 2013;46:861‐8. Baechler EC et al. Genes Immun 2004;5:347‐53. El Messaoudi S et al. Clinica Chimica Acta 2013;424:222‐30. Ahmed FE. Analytical Methods 2011;3:1029‐38. Metanomics Health – a BASF Group Company 6/5/2014