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Assessing red blood cell product quality with 2,3-DPG, ATP and p50 assays: A BEST Collaborative study
Tamar P Feldman1, Mackenzie Brandon-Coatham2,3, Jayme Kurach2,3
1American Red Cross, Holland Lab for the Biomedical Sciences, Rockville, Maryland, USA.
Vox Sanguinis
|September 23, 2025
Summary
The discontinuation of 2,3-diphosphoglycerate (2,3-DPG) assays necessitates new red blood cell (RBC) quality tests. Adenosine-5'-triphosphate (ATP) and p50 do not reliably replace 2,3-DPG for assessing RBC function.
Area of Science:
- Blood banking and transfusion medicine
- Biochemistry and metabolic analysis
- Red blood cell physiology
Background:
- Regulatory compliance mandates laboratory testing for red blood cell (RBC) quality.
- 2,3-diphosphoglycerate (2,3-DPG) has been used as a surrogate marker for RBC in vivo function, but its clinical efficacy predictive value is uncertain.
- The only validated commercial 2,3-DPG assay has been discontinued, creating a need for alternative quality assessment methods.
Purpose of the Study:
- To evaluate adenosine-5 -triphosphate (ATP) and p50 as potential replacements for 2,3-diphosphoglycerate (2,3-DPG) in assessing red blood cell quality.
- To determine the correlation between 2,3-DPG, ATP, and p50 using existing literature and retrospective data.
- To identify the need for a new assay to comprehensively characterize novel red cell products and storage conditions.
Main Methods:
- Literature review on 2,3-DPG, ATP, and p50 assays for red blood cell quality.
- Retrospective analysis of datasets from two North American blood centers.
- Evaluation of correlations between 2,3-DPG, ATP, and p50, considering biological and technical factors.
Main Results:
- Literature review did not provide sufficient evidence to support p50 as a replacement for 2,3-DPG.
- While complementary, biological and technical factors limit the correlation between 2,3-DPG and p50.
- ATP showed poor correlation with both 2,3-DPG and p50 in quality monitoring datasets and with in vivo circulation kinetics.
Conclusions:
- Existing alternative markers, ATP and p50, are not adequate replacements for 2,3-DPG in assessing red blood cell quality.
- There is a critical need for a validated replacement assay for 2,3-DPG to ensure comprehensive characterization of red blood cell products.
- Accurate assessment of red blood cell quality is essential for manufacturing, storage, and delivery, impacting clinical efficacy.
