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GingisREX: A complimentary enzyme for the detection of bacterial proteins

ABSTRACT: 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.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Escherichia Coli

SUBMITTER: Thomas Powell

LAB HEAD: Thomas John Powell

PROVIDER: PXD056345 | Pride | 2025-05-07

REPOSITORIES: Pride

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			Action	DRS
	BiotherapeuticA_GingisPrepA.raw	Raw
	BiotherapeuticA_GingisPrepB.raw	Raw
	BiotherapeuticA_GingisPrepC.raw	Raw
	BiotherapeuticA_trypsinPrepA.raw	Raw
	BiotherapeuticA_trypsinPrepB.raw	Raw

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Publications

GingisREX: A Complementary Enzyme for the Detection of Bacterial Proteins.

Powell Thomas T Widdowson Philip P Nägeli Andreas A Ebner Martin M Creese Andrew A

Journal of the American Society for Mass Spectrometry 20241011 12

Reliable enzymatic digestion underscores successful mass-spectrometry-based proteomics experiments. In this study, we compare the use of the arginine-specific protease, GingisREX, against a more traditional approach in the identification of <i>Escherichia coli</i> proteins. An increased number of protein identifications were noted when GingisREX was used compared to a trypsin/lys-C mixture. This improvement was attributed to the generation of fewer peptides per protein, resulting in a simpler pe ...[more]

PMID: 39392299

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