Non-pathogenic trypanosomatid protozoa as a platform for protein research and production

Reinhard Breitling, Susanne Klingner, Nico Callewaert, Regina Pietrucha, Anett Geyer, Gunter Ehrlich, Regina Hartung, Angelika Müller, Roland Contreras, Stephen M. Beverley, Kirill Alexandrov

Research output: Contribution to journalArticlepeer-review

128 Scopus citations

Abstract

All currently existing eukaryotic protein expression systems are based on autonomous life forms. To exploit the potential practical benefits associated with parasitic organisms we have developed a new protein expression system based on Leishmania tarentolae (Trypanosomatidae), a protozoan parasite of lizards. To achieve strong transcription, the genes of interest were integrated into the small subunit ribosomal RNA gene. Expression levels obtained were up to 30 mg of recombinant protein per liter of suspension culture and increased linearly with the number of integrated gene copies. To assess the system's potential for production of post-translationally modified proteins, we have expressed human erythropoietin in L. tarentolae. The recombinant protein isolated from the culture supernatants was biologically active, natively processed at the N-terminus, and N-glycosylated. The N-glycosylation was exceptionally homogenous, with a mammalian-type biantennary oligosaccharide and the Man3GlcNAc2 core structure accounting for >90% of the glycans present. L. tarentolae is thus the first described biotechnologically useful unicellular eukaryotic organism producing biantennary fully galactosylated, core-α-1,6-fucosylated N-glycans.

Original languageEnglish
Pages (from-to)209-218
Number of pages10
JournalProtein Expression and Purification
Volume25
Issue number2
DOIs
StatePublished - 2002

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