Expression at a 20L scale and purification of the extracellular domain of the Schistosoma mansoni TSP-2 recombinant protein A vaccine candidate for human intestinal schistosomiasis
Expression at a 20L scale and purification of the extracellular domain of the Schistosoma mansoni TSP-2 recombinant protein A vaccine candidate for human intestinal schistosomiasis
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DOI：
10.4161/hv.25787
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发表时间：
2013-11-01
影响因子：
4.8
通讯作者：
Bottazzi, Maria Elena
中科院分区：
文献类型：
--
作者：
Curti, Elena;Kwityn, Clifford;Bottazzi, Maria Elena
一种针对由曼氏血吸虫引起的人类血吸虫病的新型重组蛋白疫苗正在开发中。 Sm-TSP-2 血吸虫病疫苗由 9 kDa 重组蛋白组成，对应于独特的曼氏血吸虫四跨膜蛋白的胞外结构域。在此，我们描述了Pichia Pink（TM）分泌的Sm-TSP-2重组蛋白外环的克隆和表达，20L规模发酵的工艺开发，以及两步纯化，其蛋白回收率为31%，蛋白纯度为97%。开发的工艺适用于生产纯化蛋白，用于后续配方和一期临床研究。
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 (TM) 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.