Development and application to clinical practice of a validated HPLC method for the analysis of β-glucocerebrosidase in Gaucher disease.
The main objective of our study is to develop a simple, fast and reliable method for measuring β-glucocerebrosidase activity in Gaucher patients leukocytes in clinical practice. This measurement may be a useful marker to drive dose selection and early clinical decision making of enzyme replacement therapy. We measure the enzyme activity by high-performance liquid chromatography with ultraviolet detection and 4-nitrophenyl-β-d-glucopyranoside as substrate. A cohort of eight Gaucher patients treated with enzyme replacement therapy and ten healthy controls were tested; median enzyme activity values was 20.57mU/ml (interquartile range 19.92-21.53mU/ml) in patients and mean was 24.73mU/ml (24.12-25.34mU/ml) in the reference group, which allowed the establishment of the normal range of β-glucocerebrosidase activity. The proposed method for leukocytes glucocerebrosidase activity measuring is fast, easy to use, inexpensive and reliable. Furthermore, significant differences between both populations were observed (p=0.008). This suggests that discerning between patients and healthy individuals and providing an approach to enzyme dosage optimization is feasible. This method could be considered as a decision support tool for clinical monitoring. Our study is a first approach to in depth analysis of enzyme replacement therapy and optimization of dosing therapies.
Pharmacy Service, Dr. Peset University Hospital, Valencia, Spain; Foundation for the Promotion of Healthcare and Biomedical Research in the Valencian Community (FISABIO), Valencia, Spain. Electronic address: gras_ele@gva.es.
E Gras
EG
Colomer
Pharmacy Service, Dr. Peset University Hospital, Valencia, Spain; Foundation for the Promotion of Healthcare and Biomedical Research in the Valencian Community (FISABIO), Valencia, Spain. Electronic address: martinez_margoma@gva.es.
M A Martínez
MA
Gómez
Pharmacy Service, Obispo Polanco Hospital, Teruel, Spain. Electronic address: gonzalez_alealv@gva.es.
A González
AG
Alvarez
Pharmacy Service, Dr. Peset University Hospital, Valencia, Spain; Department of Pharmacy Technology, Faculty of Pharmacy, University of Valencia, Valencia, Spain. Electronic address: climente_mon@gva.es.
M Climente
MC
Martí
Haematology Service, Dr Peset University Hospital, Valencia, Spain.
P León
PL
Moreno
Haematology Service, Dr Peset University Hospital, Valencia, Spain.
M Fernández
MF
Zarzoso
Pharmacy Service, Dr. Peset University Hospital, Valencia, Spain; Royal National Academy of Pharmacy Member, Spain.
N V
NV
Jiménez-Torres
D000293:Adolescent; D000328:Adult; D002648:Child; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005776:Gaucher Disease / Q000175:diagnosis; D005962:Glucosylceramidase / Q000737:chemistry; D007962:Leukocytes / Q000737:chemistry; D011446:Prospective Studies; D014466:Ultraviolet Rays
D005962:Glucosylceramidase
Enzyme activity; Gaucher disease; HPLC; Imiglucerase; β-Glucocerebrosidase