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Differential mobility spectrometry (DMS) and
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Biotherapeutics utilize common cell lines for production, but during the production process, proteins from the host cells can be co-purified with the therapeutic, resulting in contamination. These residual host cell proteins (HCPs) can adversely affect drug safety, so it is critical that these HCP contaminants are accurately detected and identified during biotherapeutic development.
A common technique for detecting these contaminants is the use of an ELISA assay, developed with polyclonal antibodies raised against the host cells. These assays can take up to several months to develop and are typically capable of detecting the majority of HCPs, particularly those proteins that are highly abundant or highly immunogenic. However, detection of "the majority" simply isn’t good enough to guarantee the safety of the final biotherapeutic product, as undetected proteins that are immunogenic in humans can cause a serious reaction.
As a result, orthogonal detection and monitoring techniques, such as LC-MS/MS and CESI-MS/MS are increasingly used to supplement ELISA methodologies. LC-MS/MS enables you to not only fully detect and characterize HCPs in a biologic preparation, but also get individual protein identity information directly from the acquired data, rather than simply a number for the total sum of HCA, which is a current limitation of ELISA-based methods.
SCIEX offers a powerful solution with SWATH acquisition on a TripleTOF system. The data-independent acquisition (DIA) strategy provides unbiased sampling for detection of low-level HCPs—even those that were previously unknown or did not generate an immunogenic response in animal studies.
SCIEX offers a leading solution to the challenges presented by low-level host cell proteins (HCPs), enabling safe biotherapeutic development with proven technologies that can help you boost productivity.
SWATH acquisition on a TripleTOF system is a data-independent acquisition (DIA) strategy that provides unbiased sampling of the biologic preparation for detecting low-level HCPs—even those that were previously unknown. In addition, because of the comprehensive nature of the SWATH acquisition data file, the digital record that is created can be used as an archive of the current state of a sample. Any protein contaminant concerns that emerge in the future can be tracked over time by mining the data retrospectively, without the need to save the actual sample for reanalysis.
Host cell protein impurities in biologic product preparations can be challenging to detect due to their concentrations in the sub-ppm range. Improving ionization efficiency at ultra-low nanoliter per minute flow rates with CESI-MS can significantly enhance the detection sensitivity of these low-abundance HCPs.
To effectively keep up with program timelines, you need to be able to quickly identify, quantify and track contaminant HCPs. The unbiased SWATH acquisition method achieves consistent low-level HCP detection without time-consuming and complex up-front method development or sample fractionation procedures. Once you have developed your peptide spectral library, you can run a simple LC-MS or CESI-MS method on the protease digested sample, saving you time and expense compared to more complex 2D fractionation methods.
The additional sensitivity of the TripleTOF 6600 system is another advantage, as the quantity of host cell proteins is minute in comparison to the amount of drug product. SWATH acquisition analysis is just perfect for that kind of sample. It’s a huge benefit.
– Thomas Kofoed, Chief Executive Officer, Alphalyse
If you are responsible for monitoring host cell proteins that may contaminate a drug product preparation, you need to identify low-level proteins and monitor their presence during processing to ensure they are cleared before the drug product is released. SCIEX provides a solution that enables you to dig deeper into a complex sample, with both very high- and very low-abundance analytes, to find and monitor host cell proteins with confidence and in a single injection.
If you need to identify and monitor host cell proteins, the high-sensitivity, outstanding linear dynamic range and ultra-fast acquisition capabilities of the TripleTOF 6600 system make this the ideal technology for your needs. Unbiased SWATH acquisition delivers a high-resolution MS and MS/MS spectrum for all detectable sample components, allowing you to determine protein identities of HCP contaminants. If you are working in early development, or in sample-limited environments, coupling the TripleTOF 6600 system with the ultra-low-flow CESI 8000 Plus system will maximize sensitivity, enabling detection using less than 1 µL of sample volume. For those working in late development, the TripleTOF system can be coupled with microflow or conventional high-flow LC systems to maximize assay throughput.
Discover the power of the Zeno trap with EAD fragmentation technology in this high-resolution accurate mass system.
Start detecting the previously undetectable and quantify at lower levels with impressive precision.
Improve the sensitivity of HCP detection with reduced ion suppression and improved ionization at ultra-low nanoliter-per-minute flow rates using integrated capillary electrophoresis and electrospray ionization (CESI).
ExionLC 2.0 systems are precision analytical flow LC-MS systems that deliver the reproducibility, reliability and carryover performance your workflow demands, injection to injection, batch after batch.
Proprietary variable window SWATH acquisition provides high-speed, high-resolution MS/MS data for all detectable peptides in a sample with a single injection to enable the detection of HCPs that may have been previously unknown.
Enhancing the sensitivity of peptide quantification for the targeted host cell proteins analysis
SCIEX solutions for host cell protein analysis
Ultra-sensitive host cell protein detection using CESI-MS with SWATH acquisition
Targeted HCP quantitation workflow utilizing versatile LC-MS methodologies
The optimization of host-cell protein detection using data-independent SWATH-MS
Simultaneous quantitative peptide mapping and host cell protein detection in a recombinant IgG1 monoclonal antibody preparation using data-independent acquisition
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