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A method for red blood cell biotinylation in a closed system
Author(s) -
de Back Djuna Z.,
Vlaar Richard,
Beuger Boukje,
Daal Brunette,
Lagerberg Johan,
Vlaar Alexander P.J.,
de Korte Dirk,
van Kraaij Marian,
van Bruggen Robin
Publication year - 2018
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/trf.14535
Subject(s) - biotinylation , biotin , hemoglobin , red blood cell , reagent , chromatography , chemistry , blood product , flow cytometry , biochemistry , biology , microbiology and biotechnology , medicine , pathology
BACKGROUND Several circumstances require the accurate measurement of red blood cell (RBC) survival and clearance, such as determination of posttransfusion recovery of stored RBCs to investigate the effect of new additive solutions. To this end, biotin as a marker of RBCs to track donor RBCs in the blood of the recipient has been used in many studies. However, so far only experimental, nonvalidated, biotin‐labeled red cell concentrates (RCCs) are transfused. The goal of this study was to produce a standardized biotin‐labeled RCC product in a fast, simple, and sterile manner that can be used for clinical research and for the evaluation of new blood products according to Good Practice Guidelines (GPG) for blood establishments. STUDY DESIGN AND METHODS RCC fractions were labeled with two different concentrations of biotinylation reagent in a closed system, to prevent bacterial contamination of the end product. Using flow cytometry, the reproducibility and robustness of the biotin labeling was assessed, as well as the stability of the biotin label on the (un‐)irradiated RCC fraction. Additionally, parameters such as phosphatidylserine (PS) exposure, sodium (Na), potassium (K), free hemoglobin, adenosine triphosphate (ATP), pH, and morphology were determined prior to and after biotin labeling to rule out detrimental effects of the labeling procedure on the RCC. RESULTS Our data show that RCCs can be labeled under sterile conditions in a closed system with two different biotinylation reagent concentrations, without affecting the biological activity. CONCLUSION An easy, rapid (<2 hr), and robust method was developed to manufacture biotin‐labeled RCCs for clinical research compliant to GPG.