dc.creatorNOVO, Juliana Branco
dc.creatorOLIVEIRA, Maria Leonor Sarno
dc.creatorMAGALHES, Geraldo Santana
dc.creatorMORGANTI, Ligia
dc.creatorRAW, Isaias
dc.creatorHO, Paulo Lee
dc.date.accessioned2012-10-19T23:45:34Z
dc.date.accessioned2018-07-04T15:20:47Z
dc.date.available2012-10-19T23:45:34Z
dc.date.available2018-07-04T15:20:47Z
dc.date.created2012-10-19T23:45:34Z
dc.date.issued2010
dc.identifierMOLECULAR BIOTECHNOLOGY, v.46, n.3, p.279-286, 2010
dc.identifier1073-6085
dc.identifierhttp://producao.usp.br/handle/BDPI/25143
dc.identifier10.1007/s12033-010-9303-4
dc.identifierhttp://dx.doi.org/10.1007/s12033-010-9303-4
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1621869
dc.description.abstractDeficiency of the lysosomal glucocerebrosidase (GCR) enzyme results in Gaucher`s disease, the most common inherited storage disorder. Treatment consists of enzyme replacement therapy by the administration of recombinant GCR produced in Chinese hamster ovary cells. The production of anti-GCR antibodies has already been described with placenta-derived human GCR that requires successive chromatographic procedures. Here, we report a practical and efficient method to obtain anti-GCR polyclonal antibodies against recombinant GCR produced in Escherichia coli and further purified by a single step through nickel affinity chromatography. The purified GCR was used to immunize BALB/c mice and the induction of anti-GCR antibodies was evaluated by enzyme-linked immunosorbent assay. The specificity of the antiserum was also evaluated by western blot analysis against recombinant GCR produced by COS-7 cells or against endogenous GCR of human cell lines. GCR was strongly recognized by the produced antibodies, either as cell-associated or as secreted forms. The detected molecular masses of 59-66 kDa are in accordance to the expected size for glycosylated GCR. The GCR produced in E. coli would facilitate the production of polyclonal (shown here) and monoclonal antibodies and their use in the characterization of new biosimilar recombinant GCRs coming in the near future.
dc.languageeng
dc.publisherHUMANA PRESS INC
dc.relationMolecular Biotechnology
dc.rightsCopyright HUMANA PRESS INC
dc.rightsrestrictedAccess
dc.subjectGlucocerebrosidase
dc.subjectPolyclonal antibody
dc.subjectGaucher`s disease
dc.subjectRecombinant protein expression
dc.subjectLysosomal storage disorder
dc.titleGeneration of Polyclonal Antibodies Against Recombinant Human Glucocerebrosidase Produced in Escherichia coli
dc.typeArtículos de revistas


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