On March 24, 2020, Dr. Liangliang Sun and Ph.D. candidate Xiaojing Shen from the Department of Chemistry at Michigan State University shared their research on the separation and top-down characterization of proteoforms and protein complexes, both native and denatured, using capillary electrophoresis-mass spectrometry (CEMS) and a combination of ECD and CID (“ECciD”).

Since December 2019, Dr. Sun has been an early adopter of e-MSion ExD technology. Here, Sun and Shen discuss using an e-MSion ExD cell on an Agilent 6545XT AdvanceBioTM LC/Q-TOF to obtain extensive sequence coverage of proteins smaller than 30 kDa following separation by CE. They also describe how predicting the electrophoretic mobility of proteoforms can be used to increase confidence of proteoform IDs.

To watch the webinar, click here.

The Sun group also recently presented their work with CEMS and ECD at the 68th annual ASMS conference. Check out Xiaojing Shen’s poster: Improving CZE-MS/MS for both denatured and native top-down proteomics (ThP 544).

ASMS 2020 Reboot conference attendees can view all abstracts, posters, and oral presentations through the online portal until August 31st. Afterwards, members can find conference proceedings through the ASMS website.

On June 24th, 2:30 to 3:15 EST, Dr. Ruotolo from the Department of Chemistry at University of Michigan will speak on the advances made in collision induced unfolding (CIU) methods as part of an Agilent-hosted discussion on new tools for biotherapeutic characterization.

To register for the webinar, click here.

Agilent announces the immediate availability of the ExD AQ-250​ Option, which provides a new, powerful, and easy-to-use electron-based fragmentation technology that is a field-installed enhancement for the 6545-6560 family of Q-TOFs. The e-MSion ExD AQ-250​ Option makes possible far more complete characterization of proteins and glycoproteins with unprecedented accuracy, speed and simplicity. The ExD AQ-250​ Option is comarketed with e-MSion Inc located in Corvallis, Oregon. The technology has been developed through a ten-year collaborative effort with Agilent.

The ExD AQ-250​ Option is available for both new Agilent instruments or can be retrofitted into recent existing Q-TOF mass spectrometers. The ExD AQ-250​ Option is a drop-in replacement for the Agilent collision cell that offers superior electron fragmentation in combination with collision induced dissociation (CID) to produce more reliable identification of larger peptides and proteins than previously possible. The original functionality, sensitivity, and resolution of the Q-TOF are preserved and the ExD can be automatically optimized with Agilent’s SWARM optimization.

The ExD cell is the only electron-based technology that fast enough to fragment proteins after with ion mobility separation (IMS). When installed on Agilent’s 6560 Ion Mobility Q-Tof, the ExD AQ-250​ Option gives provides unprecedented and unique abilities to probe protein conformation after separation by ion mobility.

e-MSion’s ExD technology fragments proteins in precise and predictable ways in common with electron transfer dissociation (ETD). ExD fragmentation is simpler because no trapping is needed with a negatively charged ETD reagent. The major advantage of ExD over ETD is the ability to activate analyte ions before, during and after electron capture using CID. As a result, the ExD cell produces higher coverage of native proteins up to 25-30 kDa than possible with other commercially available fragmentation techniques and with more simplicity.

Unlike ETD, the ExD technology can produce higher energy electrons to fragment across more complex ring structures to give better characterization of glycans. In addition, ExD induces side chain fragmentation that can differentiate isobaric leucine from isoleucine, and to provide better mapping disulfide chains. The ExD cell also readily distinguishes isoaspartate from aspartate and other degradation products affecting biopharmaceutical quality.

The ExD technology preserves labile post-translational modifications on large peptides and intact proteins, which allows for the simultaneous quantitation of multiple phosphorylation and glycation sites in bottom-up and middle down proteomic work flows.

The superior dissociation of native proteins by ExD with retention of post-translational modifications and side chain differentiation opens new opportunities for more complete characterization of complex proteoforms in a highly cost-effective platform.

 

Oregon State University has announced that the 16th Uppsala Conference on Electron-Based Fragmentation has been postponed until 2021. The following notice has been released by OSU:

 

Due to the growing uncertainty of the impact of COVID-19 a decision has been made to postpone this event. We regret to inform you that the 16th Uppsala Conference on Electron-Based Fragmentation, hosted at Oregon State University has been postponed until 2021. We will add further information as it develops. If you have questions please contact Conference Services at conferences@oregonstate.edu. We thank you in advance for your patience during a time of high communications volume with our team.

 

The conference was originally scheduled to take place from August 9th-12th, including an optional Oregon Coast Excursion to conclude the event. Sponsors include e-MSion, Agilent, and OSU. For more information and updates, click here to visit the official page for UPPCON 2021.