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In vivo test of humanized L-asparaginase free and nano-encapsulated in liposomes

Grant number: 19/06919-2
Support Opportunities:Scholarships abroad - Research Internship - Doctorate
Effective date (Start): September 01, 2019
Effective date (End): August 31, 2020
Field of knowledge:Engineering - Chemical Engineering - Chemical Process Industries
Principal Investigator:Adalberto Pessoa Junior
Grantee:Eduardo Krebs Kleingesinds
Supervisor: Paul Frederick Long
Host Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade de São Paulo (USP). São Paulo , SP, Brazil
Research place: King's College London, England  
Associated to the scholarship:17/20384-9 - Development of downstream process of humanized L-asparaginase and its characterization, BP.DR


The present PhD project is part of a thematic project (Fapesp process 13/08617-7) aiming at the development of innovative L-asparaginase (ASNase) proteoforms to apply in Acute Lymphoblastic Leukemia (ALL) therapy. Currently, there is no national production of this biopharmaceutical. More specifically, we will investigate the upstream and downstream process of ASNase from Erwinia chrysanthemi expressed in a glycoengineered yeast Pichia pastoris in bioreactor. This strain, called Glycoswitch®, express proteins with glycosylation patterns more similar to human glycosilation. The association of several areas such as molecular biology and nanobiotechnology can further improve the protocols in the treatment of ALL. In this context, liposomes are nanocarriers formed by the self-aggregation of phospholipids, capable of transporting hydrophilic molecules such as proteins. One hypothesis is that glycosylations mask immunogenic sites by attenuating the immune response when injected intravenously. Another hypothesis is when this less immunogenic ASNase encapsulated in liposomes occurs an increase in its half-life time. Thereby, the main goal of this internship is to evaluate the ASNase activity, half-life time and antibody titer of Balb/c mice after the administration of the humanized and wild-type ASNase proteoforms encapsulated in liposomes (1,2-dioleoyl-sn-glycero-3-phosphocholine - DOPC). The data of these experiments are going to provide a great understanding about how the immune system reacts to these innovatives proteoform compared to the current formulations available on the market.

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
SIMAS, RODRIGO G.; KREBS KLEINGESINDS, EDUARDO; PESSOA JUNIOR, ADALBERTO; LONG, PAUL F.. An improved method for simple and accurate colorimetric determination of l-asparaginase enzyme activity using Nessler's reagent. JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY, v. 96, n. 5, . (19/06919-2, 19/12295-1)

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