Human interferon B1 ser17: Coding DNA synthesis, expression, purification and characterization of bioactive recombinant protein

dc.contributorUniversidade Estadual Paulista (UNESP)
dc.creatorVillela Dr, Anne
dc.creatorRenard, Gaby
dc.creatorPalma, Mario Sergio
dc.creatorChies, Jocelei Maria
dc.creatorDalmora, Sérgio Luiz
dc.creatorBasso, Luiz Augusto
dc.creatorSantos, Diógenes Santiago
dc.date2014-05-27T11:24:47Z
dc.date2016-10-25T18:29:03Z
dc.date2014-05-27T11:24:47Z
dc.date2016-10-25T18:29:03Z
dc.date2010-09-01
dc.date.accessioned2017-04-06T01:42:46Z
dc.date.available2017-04-06T01:42:46Z
dc.identifierJournal of Microbial and Biochemical Technology, v. 2, n. 5, p. 111-117, 2010.
dc.identifier1948-5948
dc.identifierhttp://hdl.handle.net/11449/71860
dc.identifierhttp://acervodigital.unesp.br/handle/11449/71860
dc.identifier10.4172/1948-5948.1000034
dc.identifier2-s2.0-79952607627
dc.identifierhttp://dx.doi.org/10.4172/1948-5948.1000034
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/892787
dc.descriptionA protocol to produce large amounts of bioactive homogeneous human interferon β1 expressed in Escherichia coli was developed. Human interferon β1 ser17 gene was constructed, cloned and subcloned, and the recombinant protein expressed in E. coli cells. Solubilization of recombinant human interferon β1 ser17 (rhIFN-β1 ser17) was accomplished by employing a brief shift to high alkaline pH in the presence of non-ionic detergent. The recombinant protein was purifi ed by three chromatographic steps. N-terminal amino acid sequencing and mass spectrometry analysis provided experimental evidence for the identity of the recombinant protein. Reverse phase liquid chromatography demonstrated that the content of deamidates and sulphoxides was similar to a commercial standard. Size exclusion chromatography demonstrated the absence of high molecular mass aggregates and dimers. The protocol represents an effi cient and high-yield method to obtain bioactive homogeneous monomeric rhIFN-β1 ser17 protein. It may thus represent an important step towards scaling up for rhIFN-β1 ser17 large-scale production. © 2010 Villela AD, et al.
dc.languageeng
dc.relationJournal of Microbial and Biochemical Technology
dc.rightsinfo:eu-repo/semantics/openAccess
dc.subjectBiopharmaceutical
dc.subjectBiosimilar
dc.subjectEscherichia coli
dc.subjectNon-ionic detergent
dc.subjectRecombinant human interferon β1 ser17
dc.subjectRecombinant protein production
dc.subjectinterferon beta serine
dc.subjectbacterial cell
dc.subjectbioassay
dc.subjectbiotechnological production
dc.subjectcell culture
dc.subjectcontrolled study
dc.subjectDaudi cell
dc.subjectDNA sequence
dc.subjectDNA synthesis
dc.subjectdrug screening
dc.subjectgel permeation chromatography
dc.subjectgenetic code
dc.subjecthuman
dc.subjectmass spectrometry
dc.subjectmolecular cloning
dc.subjectmolecular weight
dc.subjectnonhuman
dc.subjectnucleotide sequence
dc.subjectprotein analysis
dc.subjectprotein expression
dc.subjectprotein purification
dc.subjectrecombinant DNA technology
dc.subjectreversed phase liquid chromatography
dc.subjectsolubilization
dc.titleHuman interferon B1 ser17: Coding DNA synthesis, expression, purification and characterization of bioactive recombinant protein
dc.typeOtro


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