PAULO LEE HO
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Artigo IPEN-doc 20095 Effect of pressure on refolding of recombinant pentameric cholera toxin B2014 - RODRIGUES, D.; FARINHA-ARCIERI, L.E.; VENTURA, A.M.; CHURA-CHAMBI, R.M.; MALAVASI, R.V.; LEMKE, L.S.; GUIMARAES, J.S.; HO, P.L.; MORGANTI, L.The production of recombinant proteins is an essential tool for the expansion of modern biological research and biotechnology. The expression of heterologous proteins in Escherichia coli often results in an incomplete folding process that leads to the accumulation of inclusion bodies (IB), aggregates that hold a certain degree of native-like secondary structure. High hydrostatic pressure (HHP) impairs intermolecular hydrophobic and electrostatic interactions, leading to dissociation of aggregates under non-denaturing conditions and is therefore a useful tool to solubilize proteins for posterior refolding. Cholera toxin (CT) is composed of a non-toxic pentamer of B subunits (CTB), a useful adjuvant in vaccines, and a toxic subunit A (CTA). We studied the process of refolding of CTB using HHP. HHP was shown to be effective for dissociation of CTB monomers from IB. Posterior incubation at atmospheric pressure of concentrated CTB (1 mg/ml) is necessary for the association of the monomers. Pentameric CTB was obtained when suspensions of CTB IB were compressed at 2.4 kbar for 16 h in the presence of Tween 20 and incubated at 1 bar for 120 h. Soluble and biologically active pentameric CTB was obtained, with a yield of 213 mg CTB/liter of culture. The experience gained in this study can be important to improve the refolding of proteins with quaternary structure.Artigo IPEN-doc 12952 Natterins, a new class of proteins with kininogenase activity characterized from Thalassophryne nattereri fish venom2005 - MAGALHAES, G.S.; LOPES-FERREIRA, M.; JUNQUEIRA de AZEVEDO, I.L.M.; SPENCER, P.J.; ARAUJO, M.S.; PORTARO, F.C.V.; MA, L.; VALENTE, R.H.; JULIANO, L.; FOX, J.W.; HO, P.L.; SILVA, A.M.M. daArtigo IPEN-doc 15645 Generation of polyclonal antibodies against recombinant human glucocerebrosidase produced in Escherichia coli2010 - NOVO, JULIANA B.; OLIVEIRA, MARIA L.S.; MAGALHAES, GERALDO S.; MORGANTI, LIGIA; RAW, ISAIAS; LEE HO, PAULOArtigo IPEN-doc 18315 Generation of a chinese hamster ovary cell line producing recombinant human glucocerebrosidase2012 - NOVO, JULIANA B.; MORGANTI, LIGIA; MORO, ANA M.; LEME, ADRIANA F.P.; SERRANO, SOLANGE M. de T.; RAW, ISAIAS; LEE HO, PAULO