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Host Cell Proteome of Physcomitrella patens Harbors Proteases and Protease Inhibitors under Bioproduction Conditions
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2018-10-04 , DOI: 10.1021/acs.jproteome.8b00423 Sebastian N. W. Hoernstein 1 , Benjamin Fode 2 , Gertrud Wiedemann 1 , Daniel Lang 1, 3 , Holger Niederkrüger 2 , Birgit Berg 2 , Andreas Schaaf 2 , Thomas Frischmuth 2 , Andreas Schlosser 4 , Eva L. Decker 1 , Ralf Reski 1, 5
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2018-10-04 , DOI: 10.1021/acs.jproteome.8b00423 Sebastian N. W. Hoernstein 1 , Benjamin Fode 2 , Gertrud Wiedemann 1 , Daniel Lang 1, 3 , Holger Niederkrüger 2 , Birgit Berg 2 , Andreas Schaaf 2 , Thomas Frischmuth 2 , Andreas Schlosser 4 , Eva L. Decker 1 , Ralf Reski 1, 5
Affiliation
Host cell proteins are inevitable contaminants of biopharmaceuticals. Here, we performed detailed analyses of the host cell proteome of moss (Physcomitrella patens) bioreactor supernatants using mass spectrometry and subsequent bioinformatics analysis. Distinguishing between the apparent secretome and intracellular contaminants, a complex extracellular proteolytic network including subtilisin-like proteases, metallo-proteases, and aspartic proteases was identified. Knockout of a subtilisin-like protease affected the overall extracellular proteolytic activity. Besides proteases, also secreted protease-inhibiting proteins such as serpins were identified. Further, we confirmed predicted cleavage sites of 40 endogenous signal peptides employing an N-terminomics approach. The present data provide novel aspects to optimize both product stability of recombinant biopharmaceuticals as well as their maturation along the secretory pathway. Data are available via ProteomeXchange with identifier PXD009517.
中文翻译:
宿主细胞蛋白质组小立碗藓港口蛋白酶和下生物生产条件蛋白酶抑制剂
宿主细胞蛋白是生物药物不可避免的污染物。在这里,我们对苔藓(Physcomitrella patens)的宿主细胞蛋白质组进行了详细的分析。质谱和随后的生物信息学分析)。区分明显的分泌组和细胞内的污染物,确定了一个复杂的细胞外蛋白水解网络,包括枯草杆菌蛋白酶样蛋白酶,金属蛋白酶和天冬氨酸蛋白酶。枯草杆菌蛋白酶样蛋白酶的敲除影响了总体细胞外蛋白水解活性。除蛋白酶外,还鉴定了分泌的蛋白酶抑制蛋白,例如丝氨酸蛋白酶抑制剂。此外,我们使用N术语学方法确认了40种内源性信号肽的预期切割位点。本数据提供了新颖的方面,以优化重组生物药物的产品稳定性以及沿分泌途径的成熟度。数据可通过ProteomeXchange获得,其标识符为PXD009517。
更新日期:2018-10-04
中文翻译:
宿主细胞蛋白质组小立碗藓港口蛋白酶和下生物生产条件蛋白酶抑制剂
宿主细胞蛋白是生物药物不可避免的污染物。在这里,我们对苔藓(Physcomitrella patens)的宿主细胞蛋白质组进行了详细的分析。质谱和随后的生物信息学分析)。区分明显的分泌组和细胞内的污染物,确定了一个复杂的细胞外蛋白水解网络,包括枯草杆菌蛋白酶样蛋白酶,金属蛋白酶和天冬氨酸蛋白酶。枯草杆菌蛋白酶样蛋白酶的敲除影响了总体细胞外蛋白水解活性。除蛋白酶外,还鉴定了分泌的蛋白酶抑制蛋白,例如丝氨酸蛋白酶抑制剂。此外,我们使用N术语学方法确认了40种内源性信号肽的预期切割位点。本数据提供了新颖的方面,以优化重组生物药物的产品稳定性以及沿分泌途径的成熟度。数据可通过ProteomeXchange获得,其标识符为PXD009517。