Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 1998;12(5):263-7.
doi: 10.1002/(SICI)1098-2825(1998)12:5<263::AID-JCLA2>3.0.CO;2-7.

An enzyme-linked immunosorbent assay and reference ranges for bisphosphoglycerate mutase in human erythrocytes

T Takubo  1 , I Tsuda, N Tatsumi, K Kasuya, Y Taniguchi, T Fujita, K Uchida, Y Matsuo, T Hayashi
Affiliations

An enzyme-linked immunosorbent assay and reference ranges for bisphosphoglycerate mutase in human erythrocytes

T Takubo et al. J Clin Lab Anal. 1998.

Abstract

We established an enzyme-linked immunosorbent assay (ELISA) system for the determination of human bisphosphoglycerate mutase (BPGM) protein content in human erythrocytes using a polyclonal anti-BPGM antibody, we determined reference ranges for BPGM protein content, synthase activity, and specific activity in human erythrocytes. We produced a recombinant human BPGM (rBPGM) by gene manipulation using E. coli and then obtained the polyclonal antibody by immunizing rabbits with purified rBPGM. The reproducibility of the ELISA was in an acceptable range with a coefficient of variation under 1.5%. The ELISA was reliable in the range of 0.1 to 10 ng/mL. The polyclonal anti-rBPGM antibody did not show any cross-reaction with recombinant human B type phosphoglycerate mutase, which is highly homologous to rBPGM. The ELISA was found to be practical for the determination of BPGM protein content in human erythrocytes. The mean BPGM protein content was 56.3+/-9.7 microg/mL in whole blood (mean+/-SD, n = 50). The ELISA can be used to examine various hematologic disorders with abnormal red cell size and cell counts, and to detect BPGM enzymopathy in human erythrocytes.

PubMed Disclaimer

References

    1. Travis SF, Martinez J, Gravin J Jr, Atwater J, Gillmer P: Study of a kindred with partial deficiency of red cell 2,3‐diphosphoglycerate mutase (2,3‐DPGM) and compensated hemolysis. Blood 51: 1107–1116, 1978. - PubMed
    1. Rosa R, Prehu M‐O, Beuzard Y, Rosa J: The first case of a complete deficiency of diphosphoglycerate mutase in human erythrocytes. J Clin Invest 62: 907–915, 1978. - PMC - PubMed
    1. Calvin MC, Blouquit Y, Garel MC, Prehu MO, Cohen‐Solal M, Rosa J, Rosa R: Human bisphosphoglycerate mutase expressed in E coli: Purification, characterization, and structure studies. Biochimie 72: 337–343, 1990. - PubMed
    1. Uchida K, Kondoh K, Matsuo Y: Recombinant M‐, B‐, and MB‐type isozymes of human phosphoglyceric acid mutase: Their large‐scale production and preparation of polyclonal antibodies specific to M‐ and B‐type isozymes. Clin Chim Acta 237: 43–58, 1995. - PubMed
    1. Uchida K, Mori K, Hayashi T, Matsuo Y: Preparation of recombinant human bisphosphoglyceric acid mutase and its application. Jpn J Clin Chem 24 (Suppl 3): 75c, 1995.

Substances

LinkOut - more resources