Dataset Information

Current good manufacturing practices-compliant manufacture and measurement of biotin-labeled red blood cells.

ABSTRACT:

Background

Red blood cells (RBCs) can be labeled with N-hydroxysuccinimidobiotin (sulfo-NHS-biotin), which binds to cell surface proteins under aqueous conditions. Biotinylated RBCs can be safely infused and detected in peripheral blood samples using flow cytometry, using a fluorochrome-conjugated streptavidin (SA) detection reagent. Biotinylated RBCs have been used to track survival of transfused RBCs, and have applications in optimizing RBC storage and in understanding donor genetic, environmental and disease factors affecting RBC products.

Methods

We have developed a closed-system, current good manufacturing practices (cGMP)-compliant procedure for biotinylation of RBCs and a quantitative flow cytometric assay to estimate the dose of cell-bound biotin delivered to the patient. Resulting products were characterized for variability, sterility, endotoxin, hemolysis, total dose of cell-bound biotin and stability.

Results

The density of biotin-labeling increased as a log-linear function of sulfo-NHS-biotin-labeling concentration, with greater variability at lower concentrations. The upper estimates of biotin doses in the average product (mean RBC content = 5.55 × 10¹¹) were 9.8 and 73.0 µg for products labeled at 3 and 15 µg sulfo-NHS-biotin/mL of total reaction mixture (27 and 135 nmol/mL packed RBCs), respectively. All products were negative for bacterial and fungal growth at 14 days and were below the limit of endotoxin detection. Biotinylated RBCs were stable in vitro for up to 50 days after labeling.

Discussion

We have validated a closed-system procedure for biotinylating RBCs for investigational use. A standard operating procedure is presented in sufficient detail for implementation in a cGMP-compliant cell-processing facility.

SUBMITTER: Donnenberg AD

PROVIDER: S-EPMC10022648 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature

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Publications

Current good manufacturing practices-compliant manufacture and measurement of biotin-labeled red blood cells.

Donnenberg Albert D AD Kanias Tamir T Triulzi Darrell J DJ Dennis Catherine J CJ Moore Linda R LR Meyer E Michael EM Sinchar Derek D Kiss Joseph E JE Normolle Daniel P DP Gladwin Mark T MT

Cytotherapy 20190514 7

<h4>Background</h4>Red blood cells (RBCs) can be labeled with N-hydroxysuccinimidobiotin (sulfo-NHS-biotin), which binds to cell surface proteins under aqueous conditions. Biotinylated RBCs can be safely infused and detected in peripheral blood samples using flow cytometry, using a fluorochrome-conjugated streptavidin (SA) detection reagent. Biotinylated RBCs have been used to track survival of transfused RBCs, and have applications in optimizing RBC storage and in understanding donor genetic, e ...[more]

PMID: 31097327

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Dataset Information

Current good manufacturing practices-compliant manufacture and measurement of biotin-labeled red blood cells.

Background

Methods

Results

Discussion

Publications

Current good manufacturing practices-compliant manufacture and measurement of biotin-labeled red blood cells.

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