The Application of Automated Frozen Aliquotting Technology
The Application of Automated Frozen Aliquotting Technology Bioanalysis of Labile Compounds
Glenn Smith & Joseph Fraone CryoXtract Instruments, LLC 5 ...
The Application of Automated Frozen Aliquotting Technology Bioanalysis of Labile Compounds
Glenn Smith & Joseph Fraone CryoXtract Instruments, LLC 5 Constitution Way, Woburn MA 01801 www.cryoxtract.com
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The Application of Automated Frozen Aliquotting Technology to the Bioanalysis of Labile Compounds
Technology overview Application areas Applications within Quantitative Bioanalysis Future plans to expand bioanalytical applications Conclusions Some Things Are Better Kept Frozen!
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The Application of Automated Frozen Aliquotting Technology to the Bioanalysis of Labile Compounds
History – Frozen core sampling has long been used by geologists. – Biological sample preservation via freezing is well established and understood. – Scientists at Harvard and North-Eastern University are credited with the innovative use of analytical scale ‘Frozen Core Aliquotting’ for the purpose of preserving biospecimen integrity.
CryoXtract was established ~ 5 years ago to commercialize the use of frozen core aliquotting in various areas of life sciences and life science research. They are a subsidiary of Allied Minds, Inc., a Boston-based private investment corporation.
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The Application of Automated Frozen Aliquotting Technology to the Bioanalysis of Labile Compounds
CXT353 - compact benchtop design – semi-automated instrument – user-friendly touch screen operation – frozen aliquotting of a wide range of sample types, including: • tissues & tissue homogenates • feces • serum, plasma, whole blood, urine, and other biofluids. – Working temperature of -80°C to -150°C during aliquotting via LN2 chilling. – Eliminates damaging freeze-thaw cycles.
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The Application of Automated Frozen Aliquotting Technology to the Bioanalysis of Labile Compounds
CXT750 – automated workstation – – – – – –
Integrated touch screen operation Re-usable 3mm id coring probe Dual solvent & forced air washing station. PNP tube gripper on gantry arm Automated de-capping and re-capping Vision system automatically adjusts X/Y coring position for each core aliquot. – Up to 5 aliquots from standard 2 mL cryovials. – Aliquots delivered to tubes or plates. – Newly added ‘level detection’ feature allows for quantitative coring in ‘targeted volume’ aliquotting mode.
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The Application of Automated Frozen Aliquotting Technology to the Bioanalysis of Labile Compounds
Proprietary Coring Process
Extract 1-5 Cores From the Parent Tube
Deposit Aliquots in Single Tubes or plate wells… or Pool Together
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The Application of Automated Frozen Aliquotting Technology to the Bioanalysis of Labile Compounds
Primary Applications • BioBanking and BioRepositories. • •
RNA/DNA and all endogenous biomarkers are preserved in the primary sample. Multiple aliquots can be distributed to test labs without compromising parent sample integrity.
• Molecular Pathology. • • •
Frozen tissue coring improves tissue stability/preservation. Improved safety and ease of sampling Multi-coring enables tissue distribution investigations.
• Microbiome Research. •
Greatly simplifies the traditionally unpleasant task of sampling fecal samples.
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The Application of Automated Frozen Aliquotting Technology to the Bioanalysis of Labile Compounds Quantitative Bioanalysis – The Promise of frozen aliquotting •
Many drugs and biomarkers are unstable in biofluids such as plasma due to high levels of enzymatic activity when thawed for analysis.
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Regulators (e.g. FDA, EMA, ANVISA, etc.) continue to raise the bar for GLP assay validations, by imposing additional stability testing to better ensure assay accuracy and reproducibility upon re-analysis • •
For Biomarker research, frozen aliquotting might enable validation of lower level and/or unstable targets.
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For non-GLP discovery bioanalysis, frozen aliquotting might ensure more accurate data and better dose selection for safety/toxicity studies.
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The Application of Automated Frozen Aliquotting Technology to the Bioanalysis of Labile Compounds Priliminary Evaluation at GlaxoSmithKline (RTP, NC) 1. Evaluate general suitability of CXT750 workstation for quantitative bioanalysis • • •
Overall precision and accuracy of 50-100 uL frozen aliquotting (95% degraded after 3rd F/T
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The Application of Automated Frozen Aliquotting Technology to the Bioanalysis of Labile Compounds Ongoing work and future testing •
Preliminary evaluations within CRO & pharma analytical labs • •
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Full GLP validation / cross validation using level sensing targeted volume feature •
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Simplify pre-analytical sample collection and processing. Eliminate plasma/serum processing, and significantly reduce time from sample collection to freezing.
Uniformity of analyte distribution (homogeneity) in frozen biofluids • • • • •
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Prove that CXT-750 can be qualified for routine regulatory use in GLP bioanalysis labs
Direct frozen Whole Blood analysis. •
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Demos, remote sample exchanges, and on-site installations Generally, these are more targeted evaluations (Problem or assay specific)
Analyte considerations: small vs large molecule, protein binding Evaluate possible Radial (horizontal) and Vertical stratification. Freeze temperature considerations: -20C slow freeze, -80C freeze, flash freezing. Matrix and anti-coagulant effect: serum vs EDTA plasma vs EDTA blood. Sample volume and tube type.
Next generation platform - the CXT-’X’50
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The Application of Automated Frozen Aliquotting Technology to the Bioanalysis of Labile Compounds
Acknowledgements: GlaxoSmithKline John Dunn Chao Li Eric Yang
