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Updated: Jan 7, 2026
Rapid Characterization of Genetic Parts with Cell-Free Systems
Published on: August 30, 2021
14C-propionate incorporation assay by rapid filtration in multiwell plates
Hiroaki Kakinuma1, Azusa Kobayashi, Hiroaki Takahashi
1Department of Pediatrics, Kanazawa Medical University, 1-1 Daigaku, Uchinada, Ishikawa 920-0293, Japan. p-kaki@kanazawa-med.ac.jp
Background:
Methylmalonic aciduria (MMA) is divided into two groups according to cobalamin dependency, and this classification is important for treatment. Unfortunately, there has been no rapid and reliable method for the evaluation of cobalamin dependency.
Methods:
[14C]-propionate incorporation into intact cells in the presence of either media alone or media containing various amounts of cobalamin was measured using a 96-well filtration plate and vacuum manifold. Incorporation of radioactivity was measured by direct microplate scintillation.
Results:
Using peripheral white blood cells from normal individuals, we obtained a linear relationship between the rate of 14C-propionate incorporation and the number of cells over a broad range (10,000 to 100,000 cells/well). 14C-propionate incorporation in cells from eight patients was 1% to 13% of parallel controls.
Conclusions:
In this report, we describe a rapid, sensitive and reliable method for evaluating the cobalamin dependency of methylmalonic aciduria.
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