BLOOD SAFETY POSTER COMPILATION 2015 THERAFLEX MB-Plasma & THERAFLEX UV-Platelets

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3 BLOOD SAFETY POSTER COMPILATION 2015 THERAFLEX MB-Plasma & THERAFLEX UV-Platelets THERAFLEX MB-Plasma YEAR CONGRESS CITATION TITLE AUTHORS 2015 ISBT P ISBT P ISBT P-320 Cytomegalovirus is efficiently inactivated in human plasma by the THERAFLEX MB-Plasma system RFID tags robustness and traceability in the process of methylene blue-treated plasma THERAFLEX MB-Plasma treatment does not interfere with the antibody integrity in human plasma Handke W, Gravemann U, Lambrecht B, Sumian C, Reichenberg S, Seltsam A De Valensart N, Lotens A, Najdovski T Handke W, Gravemann U, Sumian C, Reichenberg S, Seltsam A THERAFLEX UV-Platelets YEAR CONGRESS CITATION TITLE AUTHORS 2015 ISBT P-243 Chikungunya and Ross River viruses are effectively inactivated by the THERAFLEX UV- Platelets technology Marks D, Fryk J, Prow N, Hall R, Tolksdorf F, Sumian C, Gravemann U, Seltsam A, Faddy H 2015 ISBT P-323 Effect of increased agitation speed on the in vitro quality of UV-C-treated platelet concentrates Van der Meer PF, de Laleijne-Liefting LAE, Lagerberg JW, de Korte D 3

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5 Cytomegalovirus is efficiently inactivated in human plasma by the THERAFLEX MB-Plasma system Handke W 1, Gravemann U 1, Lambrecht B 1, Sumian C 2, Reichenberg S 3, Seltsam A 1 1 Blood Center of the German Red Cross, Springe, Germany 2 Macopharma SA, Tourcoing, France 3 Macopharma International GmbH, Langen, Germany ISBT Congress 2015, London, P-301 Background Photodynamic treatment using methylene blue (MB) and visible light is in routine use for virus inactivation of human plasma. It has been shown to inactivate a broad range of different DNA and RNA viruses. Human cytomegalovirus (HCMV) is known to be transfusion transmissible and a recognized cause of morbidity and mortality in immunocompromised individuals. Aims Aim of the study was to investigate the efficacy of the THERAFLEX MB-Plasma system to inactivate HCMV in human plasma. Methods Leukodepleted plasma was prepared from whole blood using standard blood banking technology. Donors were tested for anti-cmv antibodies using the Enzygnost Anti-CMV/ IgG+IgM test to exclude the presence of neutralizing antibodies in plasma. Only anti-cmv negative plasma units were included in the study. Plasma units (n=4, 285 ml) were spiked with virus suspension (30 ml) to reach a final volume of 315 ml. MB/light treatment was done according to the manufacturer s instructions. Samples were taken after spiking (load and hold sample), after addition of MB and after illumination with different light doses. The titer of HCMV (strain AD-169, ATCC VR-538) was determined by endpoint titration and large volume plating in microtitre plate assays on MRC-5 cells (fibroblasts, ATCC CCL-171). Results The results are summarized in the Table. HCMV was efficiently inactivated by more than 4.06 log steps. Already half of the standard light dose of 120 J/cm2 resulted in inactivation of HCMV to infectivity levels below the limit of detection. THERAFLEX UV-Platelets THERAFLEX MB-Plasma SAMPLE/TIME POINT MEAN TITER [LOG 10 TCID 50 / ML] MEAN LOG 10 REDUCTION FACTOR load 4.55 ± after addition of MB 4.33 ± ± J/cm J/cm J/cm J/cm hold 4.37 ± ± 0.78 Conclusions It was shown that treatment of human plasma by THERAFLEX MB-Plasma system efficiently inactivates HCMV and thereby significantly contributes to the viral safety of plasmas for transfusion. 5

6 RFID tags robustness and traceability in the process of methylene blue-treated plasma De Valensart N, Lotens A, Najdovski T Belgian Red Cross, Namur, Belgium ISBT Congress 2015, London, P-310 Background Radio Frequency Identification (RFID) tags on bags increases possibilities of data recording during blood components processing and localisation in the storage facility. The illumination device and set for plasma Methylene Blue (MB) treatment can be equipped with RFID technology. Dedicated software is used for data transmission and management from sterile connection to illumination step of the plasma unit. Aims Evaluation of coagulation factors recovery and traceability with RFID technology during MB process. Test of tag robustness to freezing, storage and thawing in routine processing conditions. Methods Recovery experiment was performed on 12 whole blood (WB) plasmas treated with THERAFLEX MB-Plasma set and illuminated with MacoTronic B2. Samples were drawn before and after MB treatment and dosage of Fibrinogen, FVIII and Total Protein were realised (mean ± SD). RFID traceability experiment was performed on 30 bags filled with water, connected to THERAFLEX MB- Plasma set and tagged with RFID labels on the storage bag. Donation number and product code were registered in the RFID tag during the sterile connection to the treatment set. These data were transferred to MacoTrace software used to capture illumination data from MacoTronic B2. After MB treatment, bags bearing RFID tags were frozen in blast freezer (MBF42, Dometic ). Frozen units were stored at -25 C (n=20) or at -80 C (n=10) for 7 days. Frozen units were thawed either using a waterbath (n=15) or a Plasmatherm DTM (Barkey ). Robustness of RFID tags was checked through all processing steps. Process data integrity and accuracy was challenged in 10 illumination cycles that were interrupted to simulate abnormal process conditions. Finally RFID use in routine conditions was assessed with 70 WB plasmas tagged with RFID labels, treated according to routine process, frozen and stored at -25 C. Results Recoveries after MB treatment were respectively at 83 ± 10% for Fibrinogen, 87 ± 7% for FVIII and 100 ± 2% for Total Protein. Data transfers to RFID tags occurred in hidden time during sterile connection and were accurate. Donation number, product code and lot number were correctly transferred to the MacoTrace software treatment queue. MacoTronic B2 synchronised with MacoTrace database, detected tags and, if products were present in the treatment queue, automatically started illumination. Software registered process data and final status of illuminated cycle on RFID tag. The 10 interrupted cycles were detected and accurately registered on RFID tags. After handling, freezing, cold storage and thawing RFID tags (n=100) were readable. The routine treatment of 70 plasmas was fluent and no issues related to RFID tags have been detected. Conclusions RFID technology used during the THERAFLEX MB-Plasma process was functional, fast and easy to handle in routine use. In addition, RFID tags could be used for localisation in storage facility. RFID tags were readable at each processing step with an optimal traceability and data recording capability. RFID tags resisted to mechanical stress during processing, storage and in our thawing conditions. The only inconvenient is that RFID tags are not compatible with thawing devices using microwave technology. 6

7 THERAFLEX MB-Plasma treatment does not interfere with the antibody integrity in human plasma Handke W 1, Gravemann U 1, Sumian C 2, Reichenberg S 3, Seltsam A 1 1 Blood Center of the German Red Cross, Springe, Germany 2 Macopharma SA, Tourcoing, France 3 Macopharma International GmbH, Langen, Germany ISBT Congress 2015, London, P-320 Background Convalescent plasma therapy is used to treat emerging infections which cannot yet be cured by drugs or prevented by vaccination. Using plasma from patients who survived such incurable infection it is of special interest to ensure that the donated plasma is pathogen-free. THERAFLEX MB- Plasma treatment is used to inactivate pathogens in human plasma. For plasma intended to be used as convalescent plasma for treatment of acute infections, it is essential that the functionality of antibodies is preserved after the MB treatment procedure. It is known that MB/light treatment can influence the integrity of labile plasma proteins (e.g. plasma factor VIII and fibrinogen) but does not grossly reduce the function and life span of proteins in fresh frozen plasma. THERAFLEX MB-Plasma Aims The aim of the study was to investigate whether antibody binding in human plasma is affected by the THERAFLEX MB-Plasma treatment. For this purpose exemplarily the reactivity of anti-hcmv antibodies was tested prior to and after MB/light treatment. Methods Plasma units were preselected for the presence of anti-hcmv antibodies (n=4). Pathogen inactivation was done using the THERAFLEX MB-Plasma system and the MacoTronic B2 illumination device and samples were taken before and after illumination. In order to detect changes in antibody binding different dilutions of plasma samples were tested for reactivity of anti-hcmv antibodies by ELISA. THERAFLEX UV-Platelets Results Reactivity of anti-hcmv antibodies in human plasma was not reduced after MB/light treatment. Conclusions THERAFLEX MB-Plasma treatment does not negatively affect the binding of anti-hcmv antibodies in human plasma. Pathogen inactivation by MB/light treatment therefore could contribute to the safety of convalescent plasma without deteriorating its quality. 7

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9 Chikungunya and Ross River viruses are effectively inactivated by the THERAFLEX UV-Platelets technology Marks D 1, Fryk J 1, Prow N 2, Hall R 2, Tolksdorf F 3, Sumian C 4, Gravemann U 5, Seltsam A 5, Faddy H 1 1 Research and Development, Australian Red Cross Blood Service, Australia 2 School of Chemistry and Molecular Biosciences, University of Queensland, Australia 3 MacoPharma International GmbH, Langen, Germany 4 MacoPharma, Tourcoing, France 5 Blood Centre of the German Red Cross, Springe, Germany ISBT Congress 2015, London, P-243 Background Chikungunya virus (CHIKV) and Ross River virus (RRV) are mosquito-borne viruses that are of concern for transfusion safety. Given the absence of an approved blood screening test for either virus in Australia, the Australian Red Cross Blood Service restricts fresh component manufacture from individuals diagnosed with a RRV or CHIKV infection and for 4 weeks after they have recovered. In addition, similar restrictions are implemented for 4 weeks for donors returning from travel to countries at risk for CHIKV exposure. The use of pathogen reduction technology (PRT), such as the THERAFLEX UV-Platelets system, could be an alternative approach to manage the potential risk of RRV or CHIKV transfusion transmission. Aims To examine the ability of the THERAFLEX UV-Platelets system at different UVC doses to inactivate CHIKV and RRV in buffy coat-derived platelet concentrates in additive solution. Methods CHIKV or RRV were isolated and spiked into buffy coat-derived platelet concentrates in additive solution (SSP+) to give a final concentration of 6.71, or 5.30 log PFU/mL, respectively. For each virus, three buffy coat-derived platelet concentrates were spiked. Spiked platelets were then treated with the THERAFLEX UV-Platelets system at the following doses: 0.05, 0.1, 0.15 and 0.2 J/cm2 (standard dose). Pre- and post-treatment samples were taken for each dose, and the level of viral infectivity was determined using a modified version of a conventional plaque assay. The reduction in viral infectivity was calculated for each dose to determine the effect of the inactivation process. THERAFLEX MB-Plasma THERAFLEX UV-Platelets Results Treatment resulted in the inactivation of both CHIKV and RRV. Viral inactivation of 6.34 log10 for CHIKV and 5.13 log10 for RRV, at the highest UVC dose (0.2 J/cm2) was observed. A dose-dependency in viral inactivation was observed with increasing UVC doses for both viruses. Summary/Conclusions CHIKV or RRV transmission through transfusion is theoretically possible. Given this, studies are needed to gain an understanding of the threshold viral load in a blood component to cause disease through transfusion. This in turn will provide insight to the minimum level of inactivation to prevent infection through this route. Our study has shown that CHIKV and RRV, spiked into BCderived platelet concentrates in additive solution, were effectively inactivated by the THERAFLEX UV-Platelets system. At the standard dose, the residual infectivity of virus was close to, or at the limit of quantification. Our results suggest that the THERAFLEX UV-Platelets treatment could significantly contribute to the safety of platelet concentrates with respect to these arboviruses. 9

10 Effect of increased agitation speed on the in vitro quality of UV-C-treated platelet concentrates Van der Meer PF, de Laleijne-Liefting LAE, Lagerberg JW, de Korte D Sanquin Blood Bank, Department of Product and Process Development, Amsterdam ISBT Congress 2015, London, P-323 Background/Aims Pathogen reduction of platelet concentrates can take place using the Theraflex UV-Platelets procedure which is based on short-length UV-C light. However, penetration of the rays through plasma is not very high and therefore, the platelets need to be resuspended in a platelet additive solution with (±35±5% residual plasma) rather than 100% plasma. Further, the units need to be agitated vigorously during treatment to ensure that thin layers are formed where the UV-C-light can fully penetrate. The standard process uses an agitation speed of 109 rpm. Recently, studies showed that pathogen inactivation was improved by increasing the agitation speed to 180 rpm. We aimed to investigate the effect of this increased agitation speed on in vitro platelet quality. Methods Platelet concentrates from 5 pooled buffy coats in SSP+ (MacoPharma, Mouvaux, France) platelet additive solution were prepared. Three units were pooled and divided to prevent donordependent differences. One unit served as untreated control (A), one unit was agitated at 109 rpm during UV-C treatment (B), and one unit was agitated at 180 rpm during UV-C treatment, which was the investigational procedure (C). Platelet concentrates were stored for 8 days with regular measurement of in vitro quality. Results The results are summarized in the Table. The platelets show a minor increase in activation (CD62P) and apoptotic parameters (annexin A5) by day 8. Lactate production is elevated because of the UV-C-treatment, resulting in a somewhat lower ph. There was no marked difference between agitation at 109 and 180 rpm. Under all conditions, in vitro data were well within acceptance limits. Conclusions Increase of the agitation speed has minor effect on the in vitro quality of UV-C-treated platelet concentrates. Both the treated platelet units at 109 rpm and 180 rpm have excellent in vitro quality, suggesting that a higher agitation speed should not impair the clinical performance of the pathogen-reduced platelets. 10

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16 Lead the way in blood safety MACOPHARMA Rue Lorthiois F Mouvaux (France)

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