Utilize este identificador para referenciar este registo: http://hdl.handle.net/10198/9898
Título: Separation of human immunoglobulin G subclasses on a protein A monolith column
Autor: Leblebici, P.
Leblebici, M.E.
Ferreira-da-Silva, F.
Rodrigues, A.E.
Pais, L.S.
Palavras-chave: Immunoglobulin G subclasses
Protein A chromatography
Data: 2014
Citação: Leblebici, P.; Leblebici, M.E.; Ferreira-da-Silva, F.; Rodrigues, A.E.; Pais, L.S. (2014) - Separation of human immunoglobulin G subclasses on a protein A monolith column. Journal of chromatography. B. ISSN 1570-0232. 962, p. 89-93
Resumo: Monolithic columns have attracted significant attention for the purification of large biomolecules. In thepresent study, a step gradient elution method was evaluated for the separation of human immunoglobulinG (hIgG) into its subclasses on CIM (convective interaction media) r-protein A (recombinant protein A)monolithic column. hIgG was loaded onto the column and bound protein was eluted with a pH gra-dient. The subclass content of the eluted fractions was analyzed by enzyme-linked immunosorbentassay (ELISA). Results showed that separation of IgG3 from the other three subclasses can be success-fully achieved with high selectivity (100%) and throughput on monolithic media. It was also revealedthat enriched fractions of IgG1 and IgG2 could be obtained from purified hIgG in a 28 min long chro-matographic run. Three fractions with high IgG1 content (89.1%, 94.3% and 88.8%) were recovered.Furthermore, IgG2 was enriched to 64% successfully. A rapid step gradient elution scheme without anyadditives in buffers was proven to obtain enriched preparations of the two important subclasses withhigh throughput. The separation time can be reduced even more by increasing the flow rate without anyloss in selectivity, which will be beneficial in industrial scale applications.
Peer review: yes
URI: http://hdl.handle.net/10198/9898
DOI: 10.1016/j.jchromb.2014.05.029
Aparece nas colecções:CIMO - Artigos em Revistas Indexados à WoS/Scopus

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