GingisREX: A complimentary enzyme for the detection of bacterial proteins

Reliable enzymatic digestion is crucial for successful mass spectrometry-based proteomics. This study compares the arginine-specific protease, GingisREX, with a traditional trypsin/lys-C mixture for identifying E. coli proteins. Using GingisREX resulted in more protein identifications due to fewer peptides per protein, creating a simpler mixture. GingisREX also showed better digestion efficiency, fewer missed cleavages, and improved MS/MS data quality for larger peptides. These findings suggest GingisREX is a valuable complement to trypsin for detecting bacterial proteins and could be optimized as an effective alternative for identifying HCPs in biotherapeutics.

可靠的酶解操作对于基于质谱的蛋白质组学研究的成功开展至关重要。本研究针对大肠杆菌（E. coli）蛋白质的鉴定任务，对比了精氨酸特异性蛋白酶GingisREX与传统胰蛋白酶/赖氨酸内切酶C（trypsin/lys-C）混合酶解体系的性能表现。采用GingisREX进行酶解时，由于单蛋白对应肽段数量更少，样本混合物更为简洁，因此可鉴定得到更多蛋白质。此外，GingisREX还展现出更优异的酶解效率、更少的酶切漏切位点，且对大分子量肽段的串联质谱（MS/MS）数据质量具有显著提升效果。上述研究结果表明，GingisREX可作为胰蛋白酶的宝贵补充工具，用于细菌蛋白质的检测；经过优化后，其有望成为生物治疗药物中宿主细胞蛋白（Host Cell Proteins, HCPs）鉴定的高效替代方案。