Generation of a Chinese Hamster Ovary Cell Line Producing Recombinant Human Glucocerebrosidase
Joint Authors
Serrano, Solange Maria de Toledo
Moro, Ana Maria
Novo, Juliana Branco
Morganti, Ligia
Ho, Paulo Lee
Paes Leme, Adriana Franco
Raw, Isaias
Source
Journal of Biomedicine and Biotechnology
Issue
Vol. 2012, Issue 2012 (31 Dec. 2012), pp.1-10, 10 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2012-10-03
Country of Publication
Egypt
No. of Pages
10
Main Subjects
Abstract EN
Impaired activity of the lysosomal enzyme glucocerebrosidase (GCR) results in the inherited metabolic disorder known as Gaucher disease.
Current treatment consists of enzyme replacement therapy by administration of exogenous GCR.
Although effective, it is exceptionally expensive, and patients worldwide have a limited access to this medicine.
In Brazil, the public healthcare system provides the drug free of charge for all Gaucher’s patients, which reaches the order of $ 84 million per year.
However, the production of GCR by public institutions in Brazil would reduce significantly the therapy costs.
Here, we describe a robust protocol for the generation of a cell line producing recombinant human GCR.
The protein was expressed in CHO-DXB11 (dhfr−) cells after stable transfection and gene amplification with methotrexate.
As expected, glycosylated GCR was detected by immunoblotting assay both as cell-associated (~64 and 59 kDa) and secreted (63–69 kDa) form.
Analysis of subclones allowed the selection of stable CHO cells producing a secreted functional enzyme, with a calculated productivity of 5.14 pg/cell/day for the highest producer.
Although being laborious, traditional methods of screening high-producing recombinant cells may represent a valuable alternative to generate expensive biopharmaceuticals in countries with limited resources.
American Psychological Association (APA)
Novo, Juliana Branco& Morganti, Ligia& Moro, Ana Maria& Paes Leme, Adriana Franco& Serrano, Solange Maria de Toledo& Raw, Isaias…[et al.]. 2012. Generation of a Chinese Hamster Ovary Cell Line Producing Recombinant Human Glucocerebrosidase. Journal of Biomedicine and Biotechnology،Vol. 2012, no. 2012, pp.1-10.
https://search.emarefa.net/detail/BIM-505388
Modern Language Association (MLA)
Novo, Juliana Branco…[et al.]. Generation of a Chinese Hamster Ovary Cell Line Producing Recombinant Human Glucocerebrosidase. Journal of Biomedicine and Biotechnology No. 2012 (2012), pp.1-10.
https://search.emarefa.net/detail/BIM-505388
American Medical Association (AMA)
Novo, Juliana Branco& Morganti, Ligia& Moro, Ana Maria& Paes Leme, Adriana Franco& Serrano, Solange Maria de Toledo& Raw, Isaias…[et al.]. Generation of a Chinese Hamster Ovary Cell Line Producing Recombinant Human Glucocerebrosidase. Journal of Biomedicine and Biotechnology. 2012. Vol. 2012, no. 2012, pp.1-10.
https://search.emarefa.net/detail/BIM-505388
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-505388