Empowering Scientists: CIL Kits

In this educational spotlight video, we introduce QReSS™ (Quantification, Retention, and System Suitability), a novel stable isotope-labeled metabolism kit designed to enhance your MS metabolomics experiments. QReSS™ serves as a quality control solution for assessing key performance metrics, from metabolite extraction efficiency to chromatography and mass spectrometry detection. Beyond quality control, the kit’s carefully selected labeled compounds aid in metabolite identification and quantification, making QReSS™ incredibly versatile. The kit includes two dried-down stable isotope-labeled mixes and a user manual for method guidance. We demonstrate QReSS™’s utility as a spike-in standard across various sample matrices using both untargeted and targeted MS-based methods.

In this video, Robert Proos from SCIEX is joined by Dr. Andrew Percy from Cambridge Isotopes to discuss the new QReSS™ Kit for metabolomic MS initiatives. The QReSS™ Kit, developed in collaboration between SCIEX and Cambridge Isotopes, is designed to help overcome the challenges of qualifying methods and instrument platforms in mass spectrometry omics experiments. The kit consists of two dried down mixes of predominantly 13C-labeled metabolites spanning a broad metabolic class and molecular weight range. These mixes can be used as quality control checkpoints in both untargeted and targeted LC-MS metabolomic experiments for applications such as quantification, retention tracking, and system suitability. Dr. Percy highlights that unlike in-house prepared QC cocktails, the QReSS™ Kit provides a readily available and reliable solution for quality evaluations. He shares examples of how the kit has been used in matrix-free and matrix-containing conditions using a metabolomics RPLC-MRM method with dual ESI polarity. Potential applications of the QReSS™ Kit include metabolite quantification using both unlabeled and labeled mixes, with the metabolites in the mixes serving as internal standards for other metabolites in the analysis. The kit is also useful for SWATH analysis on instruments like the triple TOF. Robert Proos shares his positive experience using the QReSS™ Kit for both targeted studies with MRMs and untargeted studies on a TOF instrument, praising its wide variety of compounds.

Dr. Christoph Borchers, a leading expert in targeted proteomics, presents an in-depth technical webinar on the applications of PeptiQuant™ LC-MRM/MS Assay Kits for clinical diagnostics. He explains the fundamental principles of multiple reaction monitoring (MRM), the crucial role of PeptiQuant™ isotope-labeled internal standards, and the step-by-step process of developing highly multiplexed assays using these kits. Dr. Borchers showcases numerous examples of PeptiQuant™-based assays, demonstrating the ability to quantify hundreds of proteins in various samples such as plasma, urine, and tissues, while achieving high sensitivity and reproducibility. He shares insights into the optimization of assay conditions, the importance of rigorous validation, and the use of tools like Peptide Picker to select optimal peptides. The webinar concludes with an exciting discussion on the vast potential of dried blood spots for PeptiQuant™-based MRM analysis, highlighting how this approach could revolutionize population-wide screening by enabling less invasive, simpler, and more cost-effective sample collection.

David C. Muddiman, a distinguished professor of chemistry at North Carolina State University, introduces INLIGHT™, an innovative product from Cambridge Isotope Laboratories (CIL) for the relative quantification of N-linked glycans using mass spectrometry. The INLIGHT™ Glycan Tagging Kit contains light and heavy reagents and a multi-heptose standard, coming with a step-by-step instruction manual for easy use in complex biological systems. Potential applications are numerous, including case-control samples, pharmaceutical applications, glycan tissue imaging, cell surface glycan quantification, polysaccharide analysis, and glyco-profiling in bacteria and other species. The presentation focuses on the analysis of entire N-linked glycans in case and control samples related to cancer. A single case-control experimental setup costs around $150, while longitudinal samples (e.g., blood samples from patients over time) would require 6 reagent vials at a cost of about $650 for approximately 25 LCMS runs. Notably, these costs are all-inclusive of INLIGHT™ reagents, sample prep materials, and cleanup. Glycosylation is a crucial post-translational modification controlling cellular interactions, protein function, and protein recognition. Aberrant glycosylation has been linked to cancer since the late 1970s, underscoring its importance in understanding the disease biology. The INLIGHT™ reagents are multifunctional, tunable hydrazides. A reactive hydrazide group enables greater than 95% derivatization efficiency in less than 4 hours. A hydrophobic extension region increases glycan ionization, providing an average 4-fold increase in glycan abundance and compatibility with reverse-phase chromatography. A fennel ring allows for stable isotope incorporation (e.g., 6 13C) for relative quantification. Initial reagents were commercially purchased or synthesized in collaboration. The current reagent (fennel-GPN) provides a 4-fold improvement in ion abundance compared to free glycans, with potential to further increase hydrophobicity with larger non-porous surface areas. The methodology involves individually labeling cancer samples and control samples with light and heavy INLIGHT reagents, with normalization accounting for differences in glycan loading between individuals. The presentation will include examples of glycosylation variability in healthy individuals and details on cleaving glycans from proteomes of interest.