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

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• 作者：E. Gras Colomer ; M.A. Mart铆nez G贸mez ; A. Gonz谩lez 脕lvarez ; M. Climente Mart铆 ; P. Le贸n Moreno ; M. Fern谩ndez Zarzoso ; N.V. Jim茅nez-Torres
• 关键词：GD ; Gaucher disease ; GBA ; 尾-glucocerebrosidase ; GlcCer ; glucocerebroside ; FACS ; fluorescence-activated cell sorter ; PNP-Glc ; 4-nitrophenyl-尾-d-glucopyranoside ; PNP ; p-nitrophenol ; ERT ; enzyme replacement therapy ; IMG ; imiglucerase ; EMA ; European Medicines Agency ; HPLC&ndash ; UV ; high performance liquid chromatography with ultraviolet detection ; Tris ; Tris-(hydroxymethyl)-aminomethane ; EDTA ; ethylenediaminetraacetic acid ; ACN ; acetonitrile ; U ; unit ; MLR ; Multiple linear regression ; FDA ; Food dru
• 刊名：Journal of Pharmaceutical and Biomedical Analysis
• 出版年：25 March, 2014
• 年：2014
• 卷：91
• 期：Complete
• 页码：123-130
• 全文大小：717 K

文摘

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