Introduction

Hantaviruses belong to the family Bunyaviridae, they are enveloped RNA viruses, each carried by a specific rodent species, their natural hosts. Additionally, hantaviruses can be transmitted from rodents to humans and cause a disease. European hantaviruses are responsible for a haemorrhagic fever with renal syndrome (HFRS), but Puumala hantavirus infection has a specific name - nephropathia epidemica (NE) [1, 2, 3].

Treatment, diagnostics and vaccines for humans against hantaviruses are still at development stage in Europe. Previous studies have demonstrated, that a recombinant hantavirus nucleocapsid (N) protein derived from yeast or Escherichia coli is a suitable vaccine candidate and an antigen for diagnostics [4, 5].

Aim of our work

The aim of our work group is the development and optimisation of a process for the production of recombinant hantavirus nucleocapsid proteins (N) in the yeast Saccharomyces cerevisiae FH4C (figure 1). Additionally, we investigate downstream processing methods to improve recovery and purity of the target protein for use as a human vaccine and for diagnostics. We focus on: (a) Cultivation and scale-up of recombinant S. cerevisiae cells using batch and fed-batch cultivation strategies [7, 8]; (b) Heterologous N protein production in yeast cells; (c) Downstream processing to develop an antigen for human vaccines and diagnostics [4, 5, 6].
As a model protein we use the N protein coupled to the green fluorescent protein (6his-N-GFP) for monitoring

Figure 1. Process scheme.
A: cultivation strategy in a bioreactor
B: recombinant S. cerevisiae under light microscope
C: recombinant S. cerevisiae under fluorescence microscope (6his-N-GFP fluorescence).

Further directions

Purification of the recombinant hantavirus N protein with ion-exchange and size exclusion chromatography.

References

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