Description
Trypsin-ultra, Mass Spectrometry Grade (Recombinant) is a serine endopeptidase. It selectively cleaves peptide bonds C-terminal to lysine and arginine residues. To minimize autolysis, Trypsin-ultra, Mass Spectrometry Grade (Recombinant) has been engineered and further chemically modified by methylation. Recombinant production of the enzyme ensures it is free of contaminants from animals, including chymotrypsin and other proteolytic activities. Lack of digestion artifacts from autolysis or chemotryptic-like activity eliminates the need for manual parsing and enables fully automated reporting. This enzyme is ideal for many proteomics applications and workflows, including those using PNGase F, due to its high purity, autolysis resistance, stability, and rapid digestion times.
Product Source
Trypsin sequence from Porcine, mutagenized for enhanced resistance to autoproteolysis and expressed in E. coli.
Specificity
Selectively cleaves peptide bonds C-terminal to lysine and arginine residues.
Figure 1: Trypsin-ultra, Mass Spectrometry Grade (Recombinant) shows superior resistance to autolysis, resulting in cleaner baselines free of autolytic peptides
Samples (9 µg) of Trypsin-ultra, Mass Spectrometry Grade (Recombinant), or a different recombinant Trypsin were incubated in 100 mM Tris-HCl, 10 mM CaCl2 (pH 7.2) for 30 minutes at 37°C. After incubation, samples were injected into a Thermo Scientific® Vanquish™ UHPLC system with UV/Vis detection, coupled to a Thermo Scientific Orbitrap Exploris® 480 mass spectrometer with HCD at 32 NCE setting. Data was analyzed using Protein Metrics® (PMI) Byos® software. The chromatogram shows the UV trace at 214 nm, the arrows indicate autolytic peptides confirmed by MS/MS. The peaks corresponding to the intact Trypsin are marked with an asterisk (*).
Figure 2: Absence of autolytic artifacts with Trypsin-ultra, Mass Spectrometry Grade (Recombinant) streamlines the identification of peaks in NIST reference mAb standard
Trypsin-ultra, Mass Spectrometry Grade (Recombinant) gives clean chromatograms free of artifacts, making it ideal for regulatory compliance workflows. By delivering clean baselines, sample peaks can be identified and quantified automatically with high confidence, without the need of additional annotation or analytical expertise. The Reference Material 8671 NIST mAb (Humanized IgG1) Monoclonal Antibody was digested with Trypsin-ultra, Mass Spectrometry Grade (Recombinant) or a different recombinant Trypsin. Antibody samples (90 µg) were denatured, reduced, and alkylated. After buffer exchange and cleanup they were incubated in 100 mM Tris-HCl, 10 mM CaCl2 (pH 7.2) for 30 minutes at 37°C using a trypsin:protein ratio of 1:10. After digestion, samples were injected into a Thermo Scientific Vanquish UHPLC system with UV/Vis detection, coupled to a Thermo Scientific Orbitrap Exploris 480 mass spectrometer with HCD at 32 NCE setting. Data was analyzed using Protein Metrics (PMI) Byos software. The chromatogram shows the UV trace at 214 nm, with the arrows indicating autolytic peptides confirmed by MS/MS.
Figure 3: Maximum peptide coverage after fast, 30-minute digestion with Trypsin-ultra, Mass Spectrometry Grade (Recombinant)
Trypsin-ultra, Mass Spectrometry Grade (Recombinant) gives high-confidence and maximum peptide coverage during rapid digestion workflows, enabling high-throughput, in-process analysis. The Reference Material 8671 NIST mAb (Humanized IgG1) Monoclonal (90 µg) was denatured, reduced, and alkylated. After cleanup, the antibody was digested with Trypsin-ultra, Mass Spectrometry Grade (Recombinant) using a 1:10 trypsin:protein ratio in 100 mM Tris HCl, 10 mM CaCl2 (pH 7.2) for 30 minutes at 37°C. After digestion, samples were injected into a Thermo Scientific Vanquish UHPLC system with UV/Vis detection, coupled to a Thermo Scientific Orbitrap Exploris 480 mass spectrometer with HCD at 32 NCE setting. Data was analyzed using Protein Metrics (PMI) Byos software.
Specific Activity
The specific activity is measured by the µmol of 7-amino-4-methylcoumarin released per minute per mg of enzyme from Pro-Phe-Arg-7-amido-4-methylcoumarin at 37°C in a total reaction volume of 202.5 µL.
Reconstitution
Trypsin-ultra, Mass Spectrometry Grade (Recombinant) is provided as dried solid and should be reconstituted by the addition of 1 mL of high purity water to make a working solution of 0.1 µg/µL.
- This product is related to the following categories:
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Proteases,
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Proteome Analysis,
- This product can be used in the following applications:
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Protein Analysis Tools,
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Glycobiology & Proteomics,
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Glycoprotein Analysis