Background

Recent work has demonstrated that Red Blood Cells (RBCs) can be stored under hypoxia, resulting in enhanced glycolysis, generation of adenosine triphosphate (ATP) and 2,3-diphosphogycerate, as well as increased glutaminolysis, glutathione pools and post-transfusion recovery (Transfusion 60:786-798, 2020). We aimed to investigate the feasability of the process and product quality under GMP production conditions.

Methods

A total of 17 whole blood donations (495 ml) were separated by 5000g centrifugation and subsequent buffy coat depletion of platelets and leukocytes. RBCs were suspended in PAGGSM storage solution. Units were processed using the HEMANEXT ONE® RBC processing system (HemaNext, Lexington, MA, USA) by transfer into a separate oxygen reduction bag and subsequent storage in a PVC storage bag. Hematocrit (HCT), total Hemoglobin (Hb), ATP, O₂ saturation, hemolysis were determined on days 9, 15, 23, 30 and 42 of storage.

Results

HCT and Hb remained constant during the 42d storage with mean+SD 62.3+2.4 % and 63.1+2.3 % (HCT) and 53.5+4.7 g/dL and 53.6+4.5 g/dL (Hb) on days 0 and 42, respectively. O₂ saturation was 16.6+4.9 % (day 0) to 13.9+5.0 % (day 42). Hemolysis was 0.11+0.02% on day 0 and increased to 0.25+0.07% on day 42. ATP concentration increased from 4.07+0.59 µmol/g Hb to 4.36+0.71 µmol/g Hb on day 30, and fell to 3.36+0.58 µmol/g Hb on day 42. Irradiation did not detectably influence quality parameters within 24h.

Conclusion

GMP production of Deoxygenated RBCs is feasible, results in improved ATP levels and decreased hemolysis compared with historical controls at normoxia. Criteria suffice to apply for a manufacturing license for RBC at the responsible Länder Authority. The data form a basis for subsequent clinical evaluation of deoxygenated RBCs in order to evaluate the safety and efficacy of the novel product in chronically transfused patients.

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