Capillary Electrophoresis in Diagnosis and Monitoring of Adenosine Deaminase Deficiency

Filippo Carlucci, Antonella Tabucchi, Alessandro Aiuti, Francesca Rosi, Federica Floccari, Roberto Pagani, Enrico Marinello

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Background: The diagnosis and monitoring of severe combined immunodeficiency disease (SCID) attributable to adenosine deaminase (ADA) deficiency requires measurements of ADA, purine nucleoside phosphorylase (PNP), and S-adenosyl-L-homocysteine-hydrolase (SAHH) activity and of deoxyadenosine metabolites. We developed capillary electrophoresis (CE) methods for the detection of key diagnostic metabolites and evaluation of enzyme activities. Methods: Deoxyadenosine metabolites were separated in 30 mmol/L sodium borate-10 mmol/L sodium dodecyl sulfate (pH 9.80) at 25°C on a 60-cm uncoated capillary. For determination of enzyme activities, substrate-product separation and measurements were carried out in 20 mmol/L sodium borate (pH 10.00) at 25°C on a 42-cm uncoated capillary. Results: Deoxynucleotides and deoxyadenosine were readily detectable in erythrocytes and urine, respectively. Both methods were linear in the range 2-500 μmol/L (r >0.99). Intra- and interassay CV were 0.99 for both). In erythrocytes from healthy individuals, mean (SD) ADA activity was 5619 (2584) nmol/s per liter of packed cells. In erythrocytes of SCID patients at diagnosis, ADA activity was 56.9 (48.3) nmol/s per liter of packed cells; SAHH activity was also much reduced. PNP activity was similar in patients and controls. Conclusions: CE can be used to test ADA deficiency and enables rapid assessment of ADA expression in hematopoietic cells of SCID patients during therapy.

Original languageEnglish
Pages (from-to)1830-1838
Number of pages9
JournalClinical Chemistry
Volume49
Issue number11
DOIs
Publication statusPublished - Nov 2003

Fingerprint

Capillary electrophoresis
Adenosine Deaminase
Capillary Electrophoresis
Severe Combined Immunodeficiency
Purine-Nucleoside Phosphorylase
Monitoring
Erythrocytes
Homocysteine
Hydrolases
Metabolites
Enzyme activity
Enzymes
Sodium Dodecyl Sulfate
Urine
Severe combined immunodeficiency due to adenosine deaminase deficiency
2'-deoxyadenosine
Substrates
sodium borate
Therapeutics

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Carlucci, F., Tabucchi, A., Aiuti, A., Rosi, F., Floccari, F., Pagani, R., & Marinello, E. (2003). Capillary Electrophoresis in Diagnosis and Monitoring of Adenosine Deaminase Deficiency. Clinical Chemistry, 49(11), 1830-1838. https://doi.org/10.1373/clinchem.2003.021576

Capillary Electrophoresis in Diagnosis and Monitoring of Adenosine Deaminase Deficiency. / Carlucci, Filippo; Tabucchi, Antonella; Aiuti, Alessandro; Rosi, Francesca; Floccari, Federica; Pagani, Roberto; Marinello, Enrico.

In: Clinical Chemistry, Vol. 49, No. 11, 11.2003, p. 1830-1838.

Research output: Contribution to journalArticle

Carlucci, F, Tabucchi, A, Aiuti, A, Rosi, F, Floccari, F, Pagani, R & Marinello, E 2003, 'Capillary Electrophoresis in Diagnosis and Monitoring of Adenosine Deaminase Deficiency', Clinical Chemistry, vol. 49, no. 11, pp. 1830-1838. https://doi.org/10.1373/clinchem.2003.021576
Carlucci, Filippo ; Tabucchi, Antonella ; Aiuti, Alessandro ; Rosi, Francesca ; Floccari, Federica ; Pagani, Roberto ; Marinello, Enrico. / Capillary Electrophoresis in Diagnosis and Monitoring of Adenosine Deaminase Deficiency. In: Clinical Chemistry. 2003 ; Vol. 49, No. 11. pp. 1830-1838.
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