Qualitative and Quantitative Proteomics - Novel DIA and Processing Strategies for Complex Profiling

Data independent acquisition (DIA) is a critical technique for laboratories that need to qualitatively identify metabolites, lipids and proteins in complex samples and extract high resolution quantitative information. SONAR™, a new acquisition mode from Waters, provides quantitative MS/MS data at UPLC speeds from a data independent acquisition (DIA) method, even when samples are highly complex.

In this webinar, you will learn:

• Techniques for profiling and targeted quantitation of the human skin proteome.
• About the technology of SONAR, and how it works.
• How SONAR removes ambiguity in compound assignments, increasing your confidence and delivers better library searches, increasing the reliability and accuracy of your results.

Session 1: Advanced Data Independent Analysis for Complex Proteome Profiling

Dr. Langridge will present the technology behind SONAR, the fast scanning quadrupole data independent acquisition method that collects MS/MS results from a Data Independent Acquisition (DIA) experiment. SONAR is designed for labs that need to qualitatively identify metabolites, lipids and proteins in complex samples and extract high resolution quantitative information from a single injection. See how SONAR can give you increased confidence in your research results by getting the right answers the first time, without the need for repeated experiments.

Session 2: Global Profiling and Targeted Quantitation of the Human Skin Proteome

Allergic Contact Dermatitis is a delayed-type hypersensitivity response to small chemicals occurring in two stages; initiation, where sensitization is acquired; and elicitation, characterized by an inflammatory skin reaction. Both phases are mediated by the covalent modification of skin proteins by a chemical sensitizer; however, the cellular response to such exposure and identities of these modified proteins are currently unknown.

To enhance our understanding of the mechanisms of the allergic response, we have employed a novel data independent mass spectrometry acquisition strategy SONAR, to characterize the cellular response of human skin cells after exposure to the extreme sensitizer dinitrochlorobenzene (DNCB). To achieve this, we have generated a SuperSILAC reference standard containing 4 SILAC labeled skin cell lines and further calibrated this reference using synthetic internal reference peptides and targeted SRM to improve quantitative precision. By employing the SONAR technology, we demonstrate an increase in sensitivity and specificity, leading to increased depth of sample coverage and improved protein quantitation measurements of the skin proteome.