In MICROIMS project a novel bioanalytical platform integrating microfluidics for electrospray ionization (ESI) and ion mobility mass spectrometry (IMS MS) will be designed, introduced in human brain glycolipidomics and validated for biomarker discovery. The system to be developed is based on the coupling of fully automated chip-nanoESI with IMS MS and tandem MS (MS/MS).
The research is divided into the following 4 workpackages (WP):
WP1. Interfacing chip-nanoESI IMS MS and MS/MS. System developmen/pt in negative ion mode and optimization for simple mixtures. Testing of the system performances for the analysis of ganglioside fractions isolated from human brain.
In this WP, chip-nanoESI will be coupled with IMS MS and developed for chip-ionization, separation of ions according to their mobility, followed by fragmentation and detection of molecular ions and product ions in a single run in high-throughput mode. For development of the system, simple mixtures of gangliosides will be used.
Development of chip-nanoESI IMS MS and MS/MS for highly complex mixtures. Testing of the system performances for the analysis of gangliosides from normal human brain biopsies.
In WP2 the approach will be tested on native ganglioside mixtures extracted from human adult and fetal brain biopsies. Following the data mining and interpretation, we will complete with de novo identified species our database of structures forming the human brain gangliosidome.
WP3. Validation for glycolipid biomarker discovery in brain tumors and neurdegeneration.
In WP3 the system will be tested on native ganglioside extracts from primary benign and malignant brain tumors and neurodegeneration, the final goal being the in-laboratory platform validation for glycolipid biomarker discovery.
The last step of the research will be dedicated to completing meningioma, hemangioma and glioblastoma gangliosidome database with the newly discovered structures and proposals of biomarkers.
The general objective of MICROIMS project is to design, introduce in the field of human brain glycolipidomics and in-laboratory validate for biomarker discovery of a novel and robust analytical platform integrating microfluidics for electrospray ionization and ion mobility mass spectrometry. For this purpose a high-throughput fully automated chip-nanoESI system on a NanoMate robot will be on-line coupled to IMS MS and MS/MS, the latter being performed using the highly efficient fragmentation technique: CID. The entire on-line platform will be optimized for mapping and structural analysis of gangliosides in human brain and biomarker discovery.
Due to the combined high sensitivity, ionization and separation efficiency, the proposed platform, optimized for the investigation of brain sialylated glycolipids, has a high potential to provide better insights into the complexity of structures expressed in human brain than ever reported, and, most importantly to discover and structurally characterize new, previously not even detected, ganglioside species associated to severe brain pathologies. For these reasons it is expected that, at the end of the project, the most efficient ionization system existing nowadays and the most efficient mass spectrometry system incorporating ion mobility separation at high resolution will form an integrated, powerful platform optimized and validated for screening and identification of complex mixtures of glycans/glycoconjugates extracted from human matrices and discovery of novel molecular markers.
Platforma integrata microfluidice-spectrometrie de masa cu mobilitate ionica pentru glicolipidomica creierului uman MICROIMS
Project Number: PN-III-P2-2.1-PED-2019-0799
Project Director: CS 1 Prof. Dr. Fiz. Alina-Diana Zamfir
Project Title: Platforma integrata microfluidice-spectrometrie de masa cu mobilitate ionica pentru glicolipidomica creierului uman MICROIMS
Integrated microfluidics-ion mobility mass spectrometry platform for glycolipidomics of human brain