DisulfideDetect Banner

DisulfideDetect

Disulfide Bond Analysis - Simplified

Disulfide Bond Analysis
All Cysteine View
A) Graphical representation of the native bonds of adulimumab. The size of the circle represents the relative intensity compared to all DSBs. An "N" in the circle shows it is a "native" bond.
B) Adulimumab bonds after treatment of 70 degrees C for 30 minutes. Note the DSB scrambling.

Full Integration with Existing Software

The Disulfide Detect module is part of Bruker's existing software platform including BioTools, ProteinScape and flexAnalysis as well as integrating with other applications such as Microsoft ExcelTM for easier data Analysis and reporting.

Realize the Benefits of DisulfideDetect Software

  • Automated detection and identification of DSB's - ideal for known and unknown samples
  • Reduced Analysis Times and data complexity compared with traditional approaches....only one digest is required
  • Increased Confidence in Assignments and Data Quality due to retention time correlation and Triplet Rule

Stability and homogeneity of therapeutic antibodies are of primary concern due to the effect on safety and efficacy. Undesired biochemical, structural and conformational forms can lead to a loss of potency or, in the worse-case, severe adverse side effects. As such, the evaluation of disulfide bonds is an important aspect of gaining a comprehensive understanding of the chemical structure of a protein.

 

DisulfideDetect software is a quantum leap in ease-of-use for detecting and evaluating disulfide bonds within therapeutic proteins, such as monoclonal antibodies. It provides an easy to understand, visually-based, workspace for disulfide bond analysis for both native and non-native bonds as well as inter- and intra-chain disulfide bonds within the protein/antibody.

Breaking the Rules - the new DisulfideDetect workflow

The tertiary structure of therapeutic proteins is key to their activity and stability. Due to the complexity of these proteins and the fact that they will contain multiple disulfide bonds, analysis is a challenge – often requiring several LC/MS runs with the tryptic digests from reduced and non-reduced protein and a manual comparison of these two analyses.

The DisulfideDetect workflow allows the fully automated analysis of disulfide bonds in biopharmaceuticals, based on a single digest of the unreduced protein and without prior knowledge of enzyme specificity or native disulfide bonds. The workflow provides a semi-quantitative scrambling analysis as well.

Ease-of-use of MALDI...

DisulfideDetect software uses established methods from existing LC-MALDI instruments like the UltrafleXtremeTM, so if your lab is utilizing LC-MALDI you're all set! Just add the DisulfideDetect Module to begin analyzing disulfide bonds in your biotherapeutics. No additional hardware is necessary.

 

...Established Detection Methods

Disulfide Detect uses the Triplet Rule for screening disulfide bonds. When disulfide bonds are cleavedinside the MS, signals corresponding to the DSB peptide and the cleaved DSB peptides are generated. The sum of these masses and the triplet they form (without cleavage) gives distinctive signals in the mass spectrometer.