Utilize este identificador para referenciar este registo: http://hdl.handle.net/10400.14/6420
Título: Prediction of retention time of cutinases tagged with hydrophobic peptides in hydrophobic interaction chromatography
Autor: Lienqueo, M.E.
Salazar, O.
Henriquez, K.
Calado, C.R.C.
Fonseca, L.P.
Cabral, J.M.S
Palavras-chave: Hydrophobic interaction chromatography
Protein hydrophobicity
Retention time prediction
Hydrophobic peptide tags
Data: 2007
Editora: Elsevier
Citação: LIENQUEO, M.E. [et al.] - Prediction of retention time of cutinases tagged with hydrophobic peptides in hydrophobic interaction chromatography. Journal of chromatography A [Em linha]. 1154: 1-2 (2007) 460–463. Disponível na Internet: http://www.sciencedirect.com/science/article/pii/S0021967307006152 . ISSN 0021-9673.
Resumo: Hydrophobic interaction chromatography (HIC) is an important technique for protein purification, which exploits the separation of proteins based on hydrophobic interactions between the stationary phase ligands and hydrophobic regions on the protein surface. One way of enhancing the purification efficiency by HIC is the addition of short sequences of peptide tags to the target protein by genetic engineering, which could reduce the need for extra and expensive chromatographic steps. In the present work, a methodology for predicting retention times of cutinases tagged with hydrophobic peptides in HIC is presented. Cutinase from Fusarium solani pisi fused to tryptophan–proline (WP) tags, namely (WP)2 and (WP)4, and produced in Saccharomyces cerevisiae strains, were used as model proteins. From the simulations, the methodology based on tagged hydrophobic definition proposed by Simeonidis et al. (Φtagged), associated to a quadratic model for predicting dimensionless retention times, showed small differences (RMSE < 0.022) between observed and estimated retention times. The difference between observed and calculated retention times being lower than 2.0% (RMSE < 0.022) for the two tagged cutinases at three different stationary phases, except for the case of cut (wp)2 in octyl sepharose–2M ammonium sulphate. Therefore, we consider that the proposed strategy, based on tagged surface hydrophobicity, allows prediction of acceptable retention times of cutinases tagged with hydrophobic peptides in HIC.
Peer review: yes
URI: http://hdl.handle.net/10400.14/6420
ISSN: 0021-9673
Versão do Editor: http://www.sciencedirect.com/science/article/pii/S0021967307006152
Aparece nas colecções:FE - Artigos em revistas internacionais com Arbitragem / Papers in international journals with Peer-review

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