Plug-and-Play, Sensor-based, NanoLC/ESI Technology for Proteomics and Glycomics

DESCRIPTION (provided by applicant): Conventional nano-liquid chromatography electrospray mass spectrometry (nanoLC/ESI/MS or nanoLC/MS) is a challenging technique to perform, requiring a highly proficient user, and consequently, this powerful technique is alarmingly underutilized. The difficulty in nanoLC/MS resides in the nanoLC and electrospray moieties, as mass spectrometers and database search software have become increasingly user friendly and intuitive. NanoLC electrospray systems suffer from several major limitations including: 1) system reproducibility issues created by user inabilities in column and/or sprayer set-up; 2) difficulty making nano-fluidic connections; 3) difficulty troubleshooting system malfunctions; 4) technique is very labor intensi ve, especially at start-up; 5) cannot optimize chromatography and ionization solvent independently within a single nanoLC/MS analysis. CorSolutions has designed and will develop a robust, reliable, and user friendly nano-liquid chromatography electrospray device that will address each of the limitations of conventional nanoLC electrospray systems while still maintaining state-of-the-art performance. The CorSolutions nanoLC electrospray technology consists of conventional nanoLC pumps and a nanoLC autosample r, a hardware docking unit which mounts directly in front of the mass spectrometer orifice, and a pop-in consumable device which inserts into the hardware docking unit. The pop-in insert is a sealed vessel which serves as a black-box to the user. In th e device is imbedded a nanoLC column and spray emitter, as well as multiple sensors to diagnose and troubleshoot system issues. When the consumable device is inserted into the docking hardware, the notoriously difficult nanoLC plumbing connections are auto matically made using novel plug and play technology. The system offers the user an easy, intuitive means of performing nanoLC and electrospray, without needing to be skilled in the art. The goals of the Phase I SBIR will be to first construct a prototype system. This will include designing and building plug and play compression fittings, an automated compression fitting connection mechanism, a pop-in insert containing a nanoLC column and sprayer, sensors to detect system leaks and monitor spray curren t, as well as sensor electronics. The second goal will be to evaluate the functionality of the prototype system. This will include comparing data quality between the prototype and a conventional nanoLC system, and placing the prototype in a proteomics labo ratory for real world feedback. Our long term objective is to provide a robust, reliable nanoLC electrospray device, which can be coupled to a wide variety of mass spectrometers. We see this simple, intuitive product becoming an integral part of biologic al laboratories. And most importantly the product will allow biologists to benefit from the highly informative nanoLC/MS data, enabling scientific breakthroughs and greater understanding of complex biological systems.Nano-liquid chromatography mass spectro metry (nanoLC/MS) is a powerful technique for proteomic and glycomic studies which is able to not only identify proteins and glycans from very complex mixtures, but is also able to structurally characterize differences between the components, which is part icularly important for glycosylation post-translational modifications. However, conventional nanoLC/MS is a very difficult technique to perform, requiring a highly skilled user. Therefore this powerful technique, which could enable scientific discoveries a nd greater understanding of complex biological systems, is alarmingly underutilized. CorSolutions proposes to develop an intuitive, robust, reliable, user friendly, plug and play device, which will address the challenges of conventional nanoLC/MS, dramat ically reducing the complexity of the technique and thereby allowing pervasive use of the technology in all biological laboratories.