AB SCIEX, a global leader in life science analytical technologies, today announced that a group of Ontario-based scientists, including Kyoto Prize-winning Tony Pawson, Ph.D., and Nicolas Bisson, Ph.D., at the Samuel Lunenfeld Research Institute of Mount Sinai Hospital, have developed a new approach for protein quantification based on QTRAP^® and Eksigent technologies with multiple reaction monitoring (MRM). This new approach enables scientists to assess the time-course of protein interactions associated with specific cellular responses, improving the understanding of signaling pathways implicated in cancer and other complex illnesses. The study was published online this week in the biomedical journal Nature Biotechnology.

This first-of-its-kind study highlights the unique combination of standard affinity purification (AP) with MRM, which is also known as selective reaction monitoring (SRM). Prior to this study, researchers have had little quantitative data regarding the kinetics with which protein networks assemble and dissolve to generate specific cellular responses. It has been particularly difficult to assess the role and effects of adaptor proteins that link activated receptors to their cytoplasmic effectors.

To address this challenge, the Lunenfeld collaborated with AB SCIEX scientists to utilize the AB SCIEX QTRAP 5500 System coupled to an Eksigent NanoLC Ultra + cHiPLC nanoflex liquid chromatography system to design a new approach, which the Canadian researchers used to investigate changes in protein interactions with the adaptor protein GRB2, an important hub in growth factor signaling. Large numbers of proteins, as well as the dynamics with which they react, can be measured and quantified with this new AP-SRM method, according to the researchers.

The QTRAP 5500 System combines quantitative and qualitative analysis on a single platform, integrating a triple quadrupole for quantitative analysis with a linear accelerator trap to uniquely confirm results within the same experiment. The QTRAP system delivers high sensitivity and high performance with the highest confidence. The QTRAP 5500 is ideally suited for biomarker and proteomics applications.

Nicolas Bisson, Ph.D., the study’s first author and a scientist at the Samuel Lunenfeld Research Institute of Mount Sinai Hospital, Toronto

“We set out to better understand signaling pathways and to help other biomedical researchers go further into exploring protein interactions. By focusing on key hub proteins, the new approach we developed can be extended to obtain a truly quantitative view of dynamic intracellular protein interactions and network assembly activated by various classes of extracellular signals.”

Dave Hicks, Vice President and General Manager of the Pharma and Omics Business, AB SCIEX

“AB SCIEX has had a long-standing collaboration with the Samuel Lunenfeld Research Institute, which has some of the top biomedical researchers in the world. Their trust in using our QTRAP 5500 System and Eksigent technology to develop and conduct an innovative multiple reaction monitoring-based technique for high specificity helped uncover previously unseen kinetic insight into protein interactions, which could lead to pushing the limits in disease research.”

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