Application of the LEXSY Leishmania tarentolae system as a recombinant protein expression platform: A review

Development of a biotechnology process for the production of a novel hyperglycosylated long‐acting recombinant bovine follicle‐stimulating hormone

“…The higher glycosylation content and the consequent improved pharmacokinetic parameters of LA‐rbFSH may probably account for its enhanced bioactivity. Considering that high‐mannose glycans are the predominant glycosylation structures of the scbFSH produced by Abreu et al., [ 25 ] and that L. tarentolae lacks sialyltransferases, [ 70 ] these results emphasize, once more, the importance of a correct glycosylation pattern and, specially, an adequate sialic acid content to elicit proper in vivo bioactivity. The extended half‐life combined with the improved bioactivity may enable the use of a reduced dosage and a lower frequency of administration, potentially leading to enhanced cost‐effectiveness of the production process.…”

Expression and functional characterization of chimeric recombinant bovine follicle‐stimulating hormone produced in Leishmania tarentolae

“…During the last decade, the non‐pathogenic protozoa L. tarentolae emerged as a suitable host system for the cost‐effective production of complex mammalian proteins (de Oliveira et al., 2019 ). An important aspect of our study was to evaluate whether the production of scbFSH in L. tarentolae can be scalable to a bioreactor, as a necessary step for a potential industrial application.…”

“…The selection of the expression system for recombinant bovine FSH was based on the following rationale: (i) the hormone subunits should be expressed as a single and stable polypeptide with a suitable purification tag, (ii) the recombinant hormone should harbour a secretory signal to obtain the desired N-glycosylation pattern and to facilitate its downstream processing and (iii) high production levels should be obtained. The first goal was achieved by replicating the design of a chimeric form of His-tagged bovine FSH (Colgin, 2008) (scbFSH, Figure 1A,B), which consists of the β and α subunits linked by a CTP sequence that presents several O-glycosylation sites. Although O-glycosylation by L. tarentolae is a matter of debate, with only one paper demonstrating it (Klatt et al, 2013), this linker was selected because it allows the correct complexation of both subunits without interfering with the activity of the hormone.…”

“…Then, replacing the linker between the β and α subunits of hFSH by the carboxyl-terminal peptide (CTP) from human chorionic gonadotropin (hCG)s provided a higher expression yield and a longer half-life of bioactive rFSH both in vitro and in vivo (Klein et al, 2002;Sugahara et al, 1996). Applying this approach, an almost identical chimera of single chain bovine FSH (scbFSH) was engineered and successfully expressed in CHO cells (AspenBio Pharma, named Venaxis, Inc. since 2012; patent: (Colgin, 2008)). Recently, recombinant single chain ovine (roFSH) and bovine (bscrFSH) FSH with long-lasting stimulating activity due to the incorporation of new N-glycosylation sites in the protein sequence were successfully applied to sheep and cattle (Gutiérrez-Reinoso et al, 2022;Sanderson & Martinez, 2020).…”

The non-pathogenic protozoon Leishmania tarentolae interferes with the activation of NLRP3 inflammasome in human cells: new perspectives in the control of inflammation

“…In in vitro experiments on mammalian macrophage cells, L. tarentolae displayed a limited survival time (up to 24 h), and there is no evidence for any disease caused by this parasite in humans or other mammals (7,8). Therefore, L. tarentolae has been classified as a biosafety level I microorganism (9). Based on the absence of pathogenicity in mammals, L. tarentolae has been proposed as a vaccine platform, suitable to be exploited for the production of recombinant antigens and for their delivery to antigen presenting cells (10)(11)(12)(13).…”