Development and application to clinical practice of a validated HPLC method for the analysis of beta-glucocerebrosidase in Gaucher disease

作者：Gras Colomer E; Martinez Gomez M A; Gonzalez Alvarez A; Climente Marti M; Leon Moreno P; Fernandez Zarzoso M; Jimenez Torres N V

来源：Journal of Pharmaceutical and Biomedical Analysis, 2014, 91: 123-130.

DOI：10.1016/j.jpba.2013.12.027

• Gaucher disease
• HPLC
• beta-Glucocerebrosidase
• Enzyme activity
• Imiglucerase

The main objective of our study is to develop a simple, fast and reliable method for measuring beta-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-beta-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 20.57 mU/m1 (interquartile range 19.92-21.53 mU/m1) in patients and mean was 24.73 mU/m1 (24.12-25.34 mU/m1) in the reference group, which allowed the establishment of the normal range of beta-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. 2013 Elsevier B.V.

• 出版日期2014-3-25