Towards toxin PEGylation: The example of rCollinein-1, a snake venom thrombin-like enzyme, as a PEGylated biopharmaceutical prototype
| Autor: | Jan Tytgat, Isadora Sousa de Oliveira, Agnes A.S. Takeda, Suely Vilela Sampaio, Marcos R.M. Fontes, Steve Peigneur, Eliane Candiani Arantes, Iara Aimê Cardoso, Ernesto Lopes Pinheiro-Junior, Johara Boldrini-França, Tássia Rafaella Costa |
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| Přispěvatelé: | Universidade de São Paulo (USP), University of Vila Velha, Universidade Estadual Paulista (UNESP), KU Leuven, Universidade Federal de Uberlândia (UFU) |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Male
Toxinology Cell Survival Xenopus Biochemistry Epitope law.invention Polyethylene Glycols Structural Biology law Animals Humans Amino Acid Sequence Particle Size Molecular Biology Serine protease Biological Products Mice Inbred BALB C biology Chemistry Immunogenicity Thrombin PEGylation Fibrinogen General Medicine Snake venom thrombin-like enzyme Recombinant Proteins Kinetics Biopharmaceutical Snake venom biology.protein Recombinant DNA Leukocytes Mononuclear Female Peptides Crotalus durissus collilineatus Snake Venoms |
| Zdroj: | Scopus Repositório Institucional da UNESP Universidade Estadual Paulista (UNESP) instacron:UNESP |
| Popis: | Made available in DSpace on 2022-04-28T19:44:19Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-11-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Fonds Wetenschappelijk Onderzoek Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) KU Leuven Vlaamse regering PEGylation was firstly described around 50 years ago and has been used for more than 30 years as a strategy to improve the drugability of biopharmaceuticals. However, it remains poorly employed in toxinology, even though it may be a promising strategy to empower these compounds in therapeutics. This work reports the PEGylation of rCollinein-1, a recombinant snake venom serine protease (SVSP), able to degrade fibrinogen and inhibit the hEAG1 potassium channel. We compared the functional, structural, and immunogenic properties of the non-PEGylated (rCollinein-1) and PEGylated (PEG-rCollinein-1) forms. PEG-rCollinein-1 shares similar kinetic parameters with rCollinein-1, maintaining its capability of degrading fibrinogen, but with reduced activity on hEAG1 channel. CD analysis revealed the maintenance of protein conformation after PEGylation, and thermal shift assays demonstrated similar thermostability. Both forms of the enzyme showed to be non-toxic to peripheral blood mononuclear cells (PBMC). In silico epitope prediction indicated three putative immunogenic peptides. However, immune response on mice showed PEG-rCollinein-1 was devoid of immunogenicity. PEGylation directed rCollinein-1 activity towards hemostasis control, broadening its possibilities to be employed as a defibrinogenant agent. School of Pharmaceutical Sciences of Ribeirão Preto University of São Paulo, Av. do Café s/n° University of Vila Velha, Av. Comissário José Dantas de Melo, 21, Boa Vista II Department of Biophysics and Pharmacology Institute of Biosciences São Paulo State University (UNESP) Toxicology and Pharmacology KU Leuven, O&N II Herestraat 49 - PO box 922 Institute of Biotechnology Federal University of Uberlandia Department of Biophysics and Pharmacology Institute of Biosciences São Paulo State University (UNESP) FAPESP: 2011/23236-4 FAPESP: 2015/17286-0 FAPESP: 2015/18432-0 CNPq: 302883/2017-7 CNPq: 307155/2017-0 KU Leuven: CELSA 17/047 Vlaamse regering: GOA4919N Vlaamse regering: GOC2319N Vlaamse regering: GOE7120N KU Leuven: PDM/19/164 |
| Databáze: | OpenAIRE |
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