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Use of Tunable Copolymers in Aqueous Biphasic Systems for Extractive Bioconvers...
Mendonça, Carlos M. N.; Veríssimo, Nathalia V.; Pereira, Wellison A.; Cunha, Paula M.; Vitolo, Michele; Converti, Attilio; Kurnia, Kiki AdiAqueous biphasic systems (ABSs) based on sodium polyacrylate (NaPA), ethylene oxide/propylene oxide (EO/PO) polymers, and (EO)x-(PO)y-(EO)x triblock copolymers were prepared and applied aiming at continuous fructooligosaccharide (FOS) production and separation. EO/PO hydrophilicity/hydrophobicity balance had a significant effect on ABS formation. To develop an integrated process including the continuous enzymat...
Lysine-PEGylated Cytochrome C with Enhanced Shelf-Life Stability
Santos, João H. P. M.; Feitosa, Valker A.; Meneguetti, Giovanna P.; Carretero, Gustavo; Coutinho, João A. P.; Ventura, Sónia P. M.Cytochrome c (Cyt-c), a small mitochondrial electron transport heme protein, has been employed in bioelectrochemical and therapeutic applications. However, its potential as both a biosensor and anticancer drug is significantly impaired due to poor long-term and thermal stability. To overcome these drawbacks, we developed a site-specific PEGylation protocol for Cyt-c. The PEG derivative used was a 5 kDa mPEG-NHS...
Building better biobetters: from fundamentals to industrial application
Torres-Obreque, Karin M.; Meneguetti, Giovanna P.; Muso-Cachumba, Jorge J.; Feitosa, Valker A.; Santos, João H. P. M.; Ventura, Sónia P. M.Biological drugs or biopharmaceuticals off patent open a large market for biosimilars and biobetters, follow-on biologics. Biobetters, in particular, are new drugs designed from existing ones with improved properties such as higher selectivity, stability, half-life and/or lower toxicity/immunogenicity. Glycosylation is one of the most used strategies to improve biological drugs, nonetheless bioconjugation is an...
Characterization of levan produced by a Paenibacillus sp. isolated from Brazili...
Mendonça, Carlos M. N.; Oliveira, Rodrigo C.; Freire, Rominne K. B.; Piazentin, Anna C. M.; Pereira, Wellison A.; Gudiña, Eduardo JoséA levan-type fructooligosaccharide was produced by a Paenibacillus strain isolated from Brazilian crude oil, the purity of which was 98.5% after precipitation with ethanol and dialysis. Characterization by FTIR, NMR spectroscopy, GC-FID and ESI-MS revealed that it is a mixture of linear (26) fructosyl polymers with average degree of polymerization (DP) of 18 and branching ratio of 20. Morphological structure an...
Purification of green fluorescent protein using fast centrifugal partition chro...
Soares, Bruna P.; Santos, João H. P. M.; Martins, Margarida; Almeida, Mafalda R.; Santos, Nathalia V.; Freire, Mara G.; Santos-Ebinuma, Valéria C.The green fluorescent protein (GFP) is a biomolecule used in many biological applications such as biomarkers and biosensors, which require high purity levels. It is usually obtained from recombinant Escherichia coli strains, which also produces other endogenous proteins, demanding multiple purification steps, and consequently, increasing the overall costs to obtain pure GFP. Simpler and cheaper purification met...
Sustainable strategy based on induced precipitation for the purification of phy...
Martins, Margarida; Soares, Bruna P.; Santos, João H. P. M.; Bharmoria, Pankaj; Torres Acosta, Mario A.; Dias, Ana C. R. V.; Coutinho, João A. P.Phycobiliproteins are fluorescent proteins mainly produced by red macroalgae and cyanobacteria. These proteins, essential to the survival of these organisms, find application in many fields of interest, from medical, pharmaceutical, and cosmetic to food and textile industries. The biggest obstacle to their use is the lack of simple environmental and economical sustainable methodologies to obtain these proteins ...
Characterization of levan produced by a Paenibacillus sp. isolated from Brazili...
Mendonça, Carlos M. N.; Oliveira, Rodrigo C.; Freire, Rominne K. B.; Piazentin, Anna C. M.; Pereira, Wellison A.; Gudiña, Eduardo J.A levan-type fructooligosaccharide was produced by a Paenibacillus strain isolated from Brazilian crude oil, the purity of which was 98.5% after precipitation with ethanol and dialysis. Characterization by FTIR, NMR spectroscopy, GC-FID and ESI-MS revealed that it is a mixture of linear β(2 → 6) fructosyl polymers with average degree of polymerization (DP) of 18 and branching ratio of 20. Morphological structur...
Imidazolium-based ionic liquids as adjuvants to form Polyethylene Glycol with S...
Santos, João H. P. M.; Martins, Margarida; Silva, Amanda R. P.; Cunha, Jheniffer R.; Rangel-Yagui, Carlota O.; Ventura, Sónia P. M.Aqueous biphasic systems (ABS) are biocompatible systems applied in the extraction of biomolecules. Despite the biocompatibility of polymers and, particularly polyethylene glycol (PEG), to form ABS, their limitation in terms of phase separation is recognized. A new approach was recently proposed based on the use of ionic liquids (ILs) as adjuvants in ABS, enlarging the polarity range of these systems. Up to now...
Controlling the L‐Asparaginase extraction and purification by the appropriate s...
Magri, Agnes; Pimenta, Marcela V.; Santos, João H. P. M.; Coutinho, João A. P.; Ventura, Sónia P. M.; Monteiro, Gisele; Yagui, Carlota O. R.BACKGROUND: L-Asparaginase (ASNase) is an important biopharmaceutical for the treatment of acute lymphoblastic leukemia (ALL); however, with some restrictions due to its high manufacturing costs. Aqueous biphasic systems (ABS) have been suggested as more economical platforms for the separation/purification of proteins, but a full understanding of the mechanisms behind the ASNase partition is still a major chall...
Continuous separation of cytochrome-c PEGylated conjugates by fast centrifugal ...
Santos, João H. P. M.; Ferreira, Ana M. C.; Almeida, Mafalda R.; Quinteiro, Paula S. G. N.; Dias, Ana C. R. V.; Coutinho, João A. P.; Freire, Mara G.Herein, the effective use of aqueous biphasic systems (ABS) in Fast Centrifugal Partition Chromatography (FCPC) for the purification of PEGylated cytochrome c conjugates is shown. High recoveries (between 88% and 100%) and purities (∼100%) were obtained. Both the unreacted cytochrome c and solvents may be recovered and reused, thus allowing the design of a sustainable process in a continuous regime for the isol...