Page 95 - 00. Complete Version - Progress Report IPEN 2014-2016

Biotechnology | Progress Report 95

A work was also carried out in collaboration tion and characterization of human prolactin
with The Agency for Agribusiness Technolo- and its analogs in a research laboratory scale.
gy of Piracicaba (SP) testing different commer- In this period, the receptor antagonist G120R-
cial preparations of equine chorionic gonado- hGH was developed. A publication in “Protein
tropin (eCG) concerning ovarian stimulation Expression and Purification” (Menezes ACSC
in cattle (Alvarez RH et al., 2016). et al., 2017) describes the synthesis and char-
acterization of G120R-hGH secreted into bac-
Pirarucu (Araipama gigas) terial periplasm and obtained with a vector
pituitary gonadotropins based on a constitutive lambda PL promoter.

This antagonist can be useful for studies aim-
Araipama gigas (pirarucu) is a giant fish na- ing at investigating the effects of a simultane-
tive to the Amazon River basin that can reach ous inhibition of GH and prolactin signaling, as
up to 2 meters in length and in weigh up to a potential anti-tumoral or anti-diabetic com-
150 Kg. This species is in danger of disappear- pound. Studies related to glycosylated hPRL
ing because of the exploitation by the fishing (G-hPRL) were published in Journal of Biotech-
industry and increasing human presence. It nology (Capone MN et al., 2015). The N-glycan
is largely used for food and extractivism and structures present in native pituitary G-hPRL
commercial breeding is still incipient. In the were determined and compared with those
last period (2011-2013), our research group present in the recombinant hormone. To ob-
has isolated, from the pituitaries, and charac- tain recombinant G-hPRL, genetically modified
terized the cDNA of the α-subunit of A. gigas Chinese hamster ovary cells (CHO) adapted to
(ag) gonadotropins (FSH and LH), carrying out growth in suspension were treated with cyclo-
also a phylogenetic analysis based on its amino heximide, thus increasing the glycosylation
acid sequence. During this period (2014-2016), site occupancy from 5.5% to 38.3%, thereby fa-
the cDNAs of ag-FSHα and ag-LHα-subunits cilitating G-hPRL purification. N-Glycan profil-
have been isolated, characterized and phylo- ing proved to be a useful and accurate method-
genetically analysed. These results, including ology also for monosaccharide determination,
a three dimensional comparative modeling of molecular mass and carbohydrate content de-
ag-FSH and ag-LH, were submitted to publica- termination for the two G-hPRL preparations,
tion and are under revision by the Journal Plos in good agreement with the values obtained
One. The biotechnological synthesis of these directly via MALDI-TOF-MS.
hormones, useful for physiology and fertility
applications, will be carried out with basis on Cellular response to
ionizing radiation
these isolated genes.
Prolactin (PRL) Ionizing radiation is a physical agent known to

induce mutation and cancer, being also used
Prolactin is a 23 kDa protein hormone secret- as a widespread therapeutic modality for can-
ed by the anterior pituitary. Its therapeutic use cer treatment. Thus, one of the challenges in
is still limited, while it is important for diag- radiobiology and oncology is to understand
nostic applications such as prolactinomas, in- how the cells respond to radio-induced stress
fertility and tumorigenesis. In the context of because of exposure to radiation and its out-
recombinant hormone production, the group comes. The group, aiming to assess radio-in-
has been developing the expression, purifica- duced cytogenetic damage/mutagenesis has