NovImmune SA, 14 Chemin des Aulx, 1228 Plan-les-Ouates, Switzerland Contact: [email protected] , [email protected] * Presenting Author Assay transfer context June 8-9, 2015 Gyrolab User Seminar, London Transfer and cross-validation of a pharmacokinetic (PK) assay on the Gyrolab™ platform across three different sites (Switzerland, UK, USA) Emilie Escoffier, Anaïs Moreau, Adeline Besson * , Sabrina Lory, Walter Ferlin, Maureen Deehan and Robert Nelson Assay format Conclusions Table 1: Summary of validation parameters of PK assay at NovImmune Assay technology Case study: Transfer of a PK assay to 3 sites www.novimmune.com During the course of a drug’s development, it is often necessary to transfer assays between laboratories. The reasons for transferring can be due to limited internal resources, regulatory compliance or logistical issues. For multi-site, global clinical trials, an assay may require validation and execution in more than one site and even on different continents. This case study describes the transfer and the cross-validation of a Gyrolab™ pharmacokinetic (PK) assay in three countries and on two continents in order to achieve the goals of a clinical trial. - Rare disease and pediatric population - Rapid turnaround of data - Multi-site global clinical study sample analysis at the right time, in the right place to satisfy data delivery requirement Ensuring success in assay transfer - Evaluate the expertise - Ensure the ability to achieve a short data delivery turnaround time - Check the technology availability - Visit the CRO site - Share skills and provide training - Share information (e.g. characterization and documentation of critical reagents) - Realistic timelines - Follow-up with e-mails, teleconference calls, face-to-face meetings - Flexibility State your expectations clearly from the start - Fast data turnaround capacity - Suitable with low sample volume - Good technical support from Gyros in case of troubleshooting More challenging to transfer compared to simple ELISA methodology due to platform expertise requirements Why Gyrolab™ platform? Overview Validation summary (Step 1) Validation successfully completed at NovImmune Assessment of ruggedness of the assay across the 3 sites? Working range of the method: 62.5-8000 ng/mL * %RE ≤ ±20% and %CV ≤ 20%, except for LLOQ and ULOQ for which %RE ≤ ±25% and %CV ≤ 25% Cross-validation at 3 sites (Step 4) Establish inter-laboratory reliability: prepare and test 30 samples Figure 2: Overview of the cross-validation at 3 sites Figure 1: Steps performed for successful transfer and validation of the PK assay at 3 sites A = Reference result (Mean result from NovImmune) B = Observed result (at the CRO site) Analysis : Relative Percentage Difference (%) = Target Criteria : ± 25% of the reference value for at least 80% of the samples tested Ruggedness assessment: Figure 3: Concentration of 30 samples tested blinded at 3 sites Figure 4: Relative percentage difference of 30 samples obtained for both CROs (UK and USA) compared to the reference site (NovImmune) 87% of the samples within target criteria Ruggedness assessment passed The PK assay was successfully validated at NovImmune (Switzerland) and at the two CRO sites in the UK and USA The assessment of ruggedness was successful, demonstrating the equivalence of the assay across the three different sites Successfully supported data delivery for clinical trial at the different bioanalytical laboratory sites Study requirement Choose the CRO matching your project Ensure good communication and partnership − Abstract Laser excitation of the AF Emission measurement – fluorescent profile Drug (therapeutic mAb) Biotinylated mouse anti-idiotype mAb Alexa Fluor (AF) mouse anti-idiotype mAb Gyrolab CD 200 nL (With streptavidin coated beads) Volume Definition Chambers Mixing Chamber Streptavidin Affinity Capture Column <24 hours Sample receipt Sample analysis Data QC check Data delivery Automated nano-liter scale Gyrolab™ platform Bridging immunoassay Sequential format Validation results Passed Passed up to 1 in 2000 No hook effect Room temperature 24 hours Freeze/Thaw Cycle 5 cycles 18 months stability at both -20°C and -80°C Short term stability Long Term Stability Parameter Matrix Effect Normal matrices Lipaemic and Haemolysed matrices Passed System suitability Overall Intra- and Inter- accuracy and precision within criteria* Dilutional Linearity Hook effect 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 -40 -20 0 20 40 % Relative difference to reference concentration CRO in the UK CRO in the USA 50 Samples spiked with different levels of drug * * * * Samples below LLOQ 1 • Validation at NovImmune (Switzerland) 2 • Transfer at the 1 st CRO (UK) • Validation at the 1 st CRO (UK) 3 • Transfer at the 2 nd CRO (USA) • Validation at the 2 nd CRO (USA) 4 • Cross-validation at 3 sites 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 0 4 000 8 000 12 000 40 000 80 000 120 000 200 000 400 000 600 000 Concentration (ng/mL) NovImmune CRO in the UK CRO in the USA * Samples below LLOQ * * * Samples