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

Abstract

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.

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, JAN 2021. (19/06919-2, 19/12295-1)

KLEINGESINDS, EDUARDO KREBS; PARIZOTTO, LETICIA DE ALMEIDA; EFFER, BRIAN; MONTEIRO, GISELE; LONG, PAUL F.; ARROYO-BERDUGO, YOANA; BEHRENDS, VOLKER; ESPOSITO, MARIA TERESA; CALLE, YOLANDA; PESSOA-JR, ADALBERTO. Downstream process and evaluation of the concomitant impact of a recombinant glycosylated L-asparaginase on leukemic cancer cells and the bone marrow tumor microenvironment. Process Biochemistry, v. 131, p. 11-pg., 2023-06-09. (19/06919-2, 13/08617-7, 15/07749-2, 18/03734-9, 17/25065-9, 17/20384-9, 18/15104-0)