Evaluation of Immobilized Metal-Ion Affinity Chromatography (IMAC) as a Technique for IgG(1) Monoclonal Antibodies Purification: The Effect of Chelating Ligand and Support

dc.creatorBresolin, ITL
dc.creatorBorsoi-Ribeiro, M
dc.creatorTamashiro, WMSC
dc.creatorAugusto, EFP
dc.creatorVijayalakshmi, MA
dc.creatorBueno, SMA
dc.date2010
dc.dateAPR
dc.date2014-11-20T04:01:52Z
dc.date2015-11-26T16:05:01Z
dc.date2014-11-20T04:01:52Z
dc.date2015-11-26T16:05:01Z
dc.date.accessioned2018-03-28T22:54:07Z
dc.date.available2018-03-28T22:54:07Z
dc.identifierApplied Biochemistry And Biotechnology. Humana Press Inc, v. 160, n. 7, n. 2148, n. 2165, 2010.
dc.identifier0273-2289
dc.identifierWOS:000275455100026
dc.identifier10.1007/s12010-009-8734-5
dc.identifierhttp://www.repositorio.unicamp.br/jspui/handle/REPOSIP/65832
dc.identifierhttp://www.repositorio.unicamp.br/handle/REPOSIP/65832
dc.identifierhttp://repositorio.unicamp.br/jspui/handle/REPOSIP/65832
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1265628
dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.descriptionConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.descriptionMonoclonal antibodies (MAbs) have been used for therapies and some analytical procedures as highly purified molecules. Many techniques have been applied and studied, focusing on monoclonal antibodies purification. In this study, an immobilized metal affinity chromatography membrane was developed and evaluated for the purification of anti-TNP IgG(1) mouse MAbs from cell culture supernatant after precipitation with a 50% saturated ammonium sulfate solution. The chelating ligands iminodiacetic acid, carboxymethylated aspartic acid (CM-Asp), nitrilotriacetic acid, and tris (carboxymethyl) ethylenediamine in agarose gels with immobilized Ni(II) and Zn(II) ions were compared for the adsorption and desorption of MAbs. The most promising chelating ligand-CM-Asp-was then coupled to poly(ethylene vinyl alcohol) (PEVA) hollow fiber membranes. According to SDS-PAGE and ELISA analyses, a higher selectivity and a purification factor of 85.9 (fraction eluted at 500 mM Tris) were obtained for IgG(1) using PEVA-CM-Asp-Zn(II). The anti-TNP MAb could be eluted under mild pH conditions causing no loss of antigen binding capacity.
dc.description160
dc.description7
dc.description2148
dc.description2165
dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.descriptionConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.descriptionConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.languageen
dc.publisherHumana Press Inc
dc.publisherTotowa
dc.publisherEUA
dc.relationApplied Biochemistry And Biotechnology
dc.relationAppl. Biochem. Biotechnol.
dc.rightsfechado
dc.sourceWeb of Science
dc.subjectMonoclonal antibodies
dc.subjectPurification
dc.subjectAdsorption
dc.subjectIMAC
dc.subjectAffinity membrane
dc.subjectDownstream processing
dc.subjectPolyacrylamide Gels
dc.subjectBuffer Systems
dc.subjectProteins
dc.subjectMembrane
dc.subjectSeparation
dc.subjectNickel
dc.subjectAdsorption
dc.subjectSuccesses
dc.subjectFragments
dc.subjectHistidine
dc.titleEvaluation of Immobilized Metal-Ion Affinity Chromatography (IMAC) as a Technique for IgG(1) Monoclonal Antibodies Purification: The Effect of Chelating Ligand and Support
dc.typeArtículos de revistas


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