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

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).…”