Analysis of glucocerebrosidase activity in dry blood spots using tandem mass spectrometry
2010; Elsevier BV; Volume: 412; Issue: 3-4 Linguagem: Inglês
10.1016/j.cca.2010.11.006
ISSN1873-3492
AutoresElisa Legini, Joseph J. Orsini, Christina Hung, Monica Martin, Amanda Showers, Maurizio Scarpa, X. Kate Zhang, Joan Keutzer, Adolf Mühl, Olaf A. Bodamer,
Tópico(s)Carbohydrate Chemistry and Synthesis
ResumoGaucher disease (GD) is due to deficiency of acid-β-glucosidase (ABG) and comprises a clinical spectrum with variable age of onset and severity. We evaluated a tandem mass spectrometry (MS/MS) method to measure ABG activity for high through-put screening.ABG activity was measured in 3.2 mm punches from dry blood spots (DBS). Each punch was incubated for 21 h with the substrate D-Glucosyl-β1-1'-N-dodecanoyl-D-erythro-sphingosine [C12-glucocerebroside (C(36)H(69)NO(8))] and internal standard N-myristoyl-D-erythro-sphingosine [C14-ceramide (C(32)H(63)NO(3))]. The product and internal standard were quantified using MS/MS.ABG activities in anonymized newborn screening samples from NY State were (mean) 22.0 μmol/h/L±(SD) 13.8 μmol/h/L (n=2088, median 19.9 μmol/h/L, 95%CI 22.59-21.41 μmol/h/L). The enzymatic activity in DBS from 10 treatment naïve adult Gaucher patients was less than 4.2 μmol/h/L. ABG activity was stable for 3 months at room temperature a 20% activity reduction was observed. Inter- and intra-run imprecisions were 8% and 13.7%, respectively. The limit of detection was 0.75 μmol/h/L and limit of quantification was 1.25 μmol/h/L.The measurement of ABG activities in DBS using MS/MS is suitable for high-throughput analysis of at-risk individuals and potentially for newborn screening for GD.
Referência(s)