GEN-MKT-18-7897-A
May 11, 2016 | Biopharma, Blogs | 0 comments
Fast LC-MS acquisition and automated data processing will help you speed up peptide mapping of your biotherapeutic, including critical disulfide bond and post-translational modification characterization. SCIEX helps you untangle the complexity of disulfide bonds, speeding up your characterization process.
See the full article by downloading the compendium >
Accurate disulfide bond mapping is essential for correctly establishing structure-function relationships as well as for monitoring the structural integrity of recombinant monoclonal antibodies (mAbs) throughout their production. Inappropriate disulfide bonds can affect a mAb’s stability, potency, aggregation, and may also signal errors in the cell culture or purification process. By following a biotherapeutic’s disulfide patterns over time, manufacturers can quickly detect production problems and then correct them as early as possible.
Correctly assigning disulfide bonds in a mAb, or ADC can be challenging and time-consuming due to the heterogeneity, large size, and multiple cysteine residues found in these biomolecules. Traditional approaches for disulfide mapping are based on fast liquid chromatography-mass spectrometry (LC-MS) analysis; however, these methods can be inefficient and usually involve digestion with multiple enzymes, tedious data processing, and intensive manual inspection of chromatograms for the identification of any possible disulfide linkages.
As the biotherapeutics industry develops and expands, there is an urgent need for software tools that can rapidly facilitate and accelerate the higher-order structural characterization of biopharmaceutical products. To meet these requirements, SCIEX has developed BioPharmaView™ Software, a data processing suite that can reduce the complexity of the massive data sets generated during biotherapeutic analysis. BioPharmaView Software uses rapid processing tools to accelerate critical characterization assays–such as peptide mapping and disulfide bond identification– by automating peak assignments, simplifying data processing, and streamlining the reporting process.
Peak Assignment Reduces Time for Peptide Mapping ExperimentsTo identify and match peptides, BioPharmaView Software automatically scores b- and y- ions from the high-resolution MS/MS spectra; and then the highest scoring experimental peaks are compared to a list of theoretical masses automatically generated by the software. The peak assignment process is further enhanced by predicting the theoretical fragment ion masses for non-reduced, disulfide-linked peptides before comparison with experimental data. Including other criteria in the ion selection process–such as MS/MS scoring, multiple charge states, and a retention time (RT) filter–can also help reduce the time needed for peptide mapping experiments. This enables manufacturers to meet regulatory requirements more quickly during the production and marketing of a new biotherapeutic product.
The ResultsIn this article, we successfully developed an efficient and automated workflow that comprehensively identified every disulfide linkage in the Fab region of an mAb. The use of the high-speed, TripleTOF® LC-MS System contributed to time-savings during disulfide analysis by permitting accurate mass MS and MS/MS information to be collected simultaneously, providing the high-resolution data necessary for differentiating closely related species and confirming structural assignments. And by using BioPharmaView Software to process the dataset, identifying the location of five disulfide linkages in the Fab region of an mAb was completed in a fast and automated fashion.
See the results in the full article by downloading the Biologics Analytical Characterization Compendium >
The Echo® MS+ system is a novel platform for Acoustic Ejection Mass Spectrometry (AEMS) and combines the speed of acoustic sampling with the selectivity of mass spectrometry. This platform has been designed for high throughput analysis of small and large molecules. The technology combines Acoustic Droplet Ejection (ADE), an Open Port Interface (OPI) and could be coupled with the SCIEX Triple Quad 6500+ system or the ZenoTOF 7600 system.
The Echo® MS+ system comprises of an open-port interface (OPI) and acoustic droplet ejection (ADE) module which could be coupled with a mass spectrometer. The mass spectrometer could either be a SCIEX Triple Quad 6500+ system or the ZenoTOF 7600 system. This non-liquid chromatography based; high-throughput screening platform enables rapid analysis of compounds at speeds of up to 1 sample/second.
The ability to consistently achieve reproducible results on many complex samples across multiple days is critical to a routine clinical laboratory. Laboratories relying on analytical instrumentation require stability and robustness to perform a variety of screening and confirmatory assays with confidence. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) has become the preferred analytical method in the clinical laboratory to reliably perform clinical testing as it provides best-in-class performance and reliability for the most challenging assays. LC-MS/MS offers the required levels of sensitivity and specificity for the detection and quantitation of molecules from complex biological samples, helping laboratories deliver highly accurate data for a variety of clinically relevant analytes across a wide range of assays.
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