Expression at a 20L scale and purification of the extracellular domain of theSchistosoma mansoniTSP-2 recombinant protein

Abstract

A novel recombinant protein vaccine for human schistosomiasis caused by Schistosoma mansoni is under development. The Sm-TSP-2 schistosomiasis vaccine is comprised of a 9 kDa recombinant protein corresponding to the extracellular domain of a unique S. mansoni tetraspanin. Here, we describe the cloning and the expression of the external loop of Sm-TSP-2 recombinant protein secreted by Pichia Pink the process development at 20L scale fermentation, and the two-steps purification, which resulted in a protein recovery yield of 31% and a protein purity of 97%. The developed processes are suitable for the production of purified protein for subsequent formulation and Phase 1 clinical studies.

Impact of Schistosoma mansoni on Malaria Transmission in Sub-Saharan Africa

“…Though our study focused on Plasmodium falciparum , our results may be applicable to other forms of malaria such as Plasmodium ovale and Plasmodium vivax , which may also interact with S. mansoni . Prototype vaccines for both malaria [34] and S. mansoni intestinal schistosomiasis [35] are under development, such that the two vaccines could be co-formulated or combined [36] . Our results suggest that a co-formulated or combined vaccine may be more efficacious in reducing malaria transmission in S. mansoni endemic communities than a vaccine targeting malaria alone.…”

Non-conventional expression systems for the production of vaccine proteins and immunotherapeutic molecules

“…Over the years, P. pastoris has become a widely used expression system, [24][25][26][27][28][29] and has been investigated for the production of recombinant protein antigens for human vaccines. 30,31 Several examples of recombinant protein antigen expression in P. pastoris show how this system can offer the benefit of higher eukaryotic host cells, with glycosylated and well-folded proteins, while having the advantage of being easy and inexpensive to cultivate.…”

“…It shows low viscosity morphology, allowing easy, large-scale, high-cell-density growth in fermenters using inexpensive substrates. Furthermore, this fungus supports broad ranges of pH (4.5-7.0) and temperature (25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39)(40)(41)(42)(43)(44) C) for growth. A C1 genetic toolbox is available, and the 3eight megabase pair genome has been sequenced, enabling the development of various gene expression strategies.…”

Process Characterization and Biophysical Analysis for a Yeast-Expressed Phlebotomus papatasi Salivary Protein (PpSP15) as a Leishmania Vaccine Candidate

“…With 79% of high overall recovery, we expect to collect approximately 785 mg of purified PpSP15 per litter of FS. When comparing with the purification process for several other reported recombinant protein vaccine antigens at similar fermentation scale, 24,27,[29][30][31][32] one can find that the process for PpSP15 requires much less volume of resins; in addition, size exclusion chromatography, a process not ideal for large-scale production, had been included in the purification process for some of these recombinant protein vaccine antigens to increase their purity. 24,27,30,31 However, the current purification scheme for PpSP15 does not require an size exclusion chromatography step to improve the purity to >95%, and all the chosen purification steps are scalable, except that dialysis was used to exchange PpSP15 into an appropriate buffer due to the small volumes at this development phase.…”