Programme
Mass Spectrometry 2012 will offer a diverse scientific programme from leading technology and training experts in mass spectrometry and MS-hyphenated systems. Below is an outline of the topics to be included in the programme:
Analytical Method Management
• LC/MS development and optimization
• GC/MS development and optimization
• Validation of qualitative methods
• Validation of quantitative methods
• Regulatory issues
Collision/Fragmentation
• In-source CAD and MS/MS for qualitative methods
• SRM/SIM for quantitation
• ECD/ETD
• New technology (T-Wave, TOF-TOF, etc.)
Interpretation and Informatics
• Use of libraries and databases for interpretation
• MS software systems: bio interpretation, isotope ratio, small molecule interpretation aids
• LIMS, ELN, regulatory considerations
Emerging Technology
• DART, DESI open air technology
• TOF technology
• Ion Trap technology
Illustrative Applications
• Pharma
• Biotech
• Forensic
• Food
• Petrochemical
• Environmental
• Other industries
Some key presentations include:
Mass Spectrometry Imaging and Ambient Mass Spectrometry in Clinical, Biological and Environmental Research
Bernhard Spengler (Justus-Liebig Universitat Giessen, Germany)
New minimum-‐preparation and in situ methods were employed in our lab that provide a closer look into the mechanisms of life. They include direct ionization techniques such as DESI, REIMS, LDI or LTP, on the one hand and atmospheric pressure MALDI methods for high resolution tissue imaging, on the other hand. They also include miniaturized mass analyzers, ion/aerosol transportation systems for field analysis, electrospray postionization methods and dedicated software tools for data evaluation. The presentation will summarize results from our research focus funded by the state of Hesse, in the field of instrumentation and methodology. For MALDI imaging of biological tissue, a versatile method providing high resolution in mass and space has been developed and applied to several research areas. The new atmospheric pressure ion source was coupled to an FTMS instrument (Orbitrap or Exactive, Thermo Fisher Scientific, Bremen), providing a spatial resolution on tissue of 3 to 5 micrometer, at a mass accuracy of 2 ppm. Ion images were acquired with a speed of more than 2 pixels per second. The presentation will include examples of imaging phospholipids, drug compounds and peptides from mammalian tissue and plant tissue. It will also describe the technical principles of the new instrumentation and its availability.
Where Are We With Respect to Using Mass Spectrometry to Identify Unknowns and Known Unknowns Using Databases and Third Party Mass Spectral Software
David Sparkman (University of the Pacific, USA)
In the past decade databases of spectra obtained by MS/MS techniques have appeared, stabilized and grown in size. Algorithms have been developed to do comparisons of operator generated spectra with these databases leading to successful identification. There have been a number of high quality application-specific databases of EI spectra coming from experts in these applications. In the area of gas chromatography/mass spectrometry (GC/MS) that other dimension is especially important to an identification where the mass spectra of different compounds are isobaric, the retention index has now been cataloged and has become computer useable as an aid in method development. Non-mass spectral databases such as ChemSpider and Chem Abstracts have also been used with information extracted from mass spectral data to aid in an identification of especially what has become know as know unknowns. Another important area is that of the role of accurate mass measurements. Now that such data are more accessible than ever before, through advances in time-of-flight (TOF) mass spectrometers and the use of the CERNO Bioscience Mass Works software with integer mass data, this along with information in the above-described databases cannot be ignored. All of these areas of the current status of databases, accurate mass, and search programs will be presented in some detail.
Top-Down Mass Spectrometry Methods for Defining Proteins and Protein Interactions Important in Biology and Medicine
Joseph Loo (UCLA Molecular Biology Institute, USA)
The application of mass spectrometry (MS) for studying proteins and protein complexes has utility in biology, biochemistry, and biomedical research. The assessment of protein interactions can address the functional role of proteins and protein complexes. Developments in protein MS and tandem MS (MS/MS) to define the structures of proteins and protein complexes, including the sites of ligand bonding will be discussed. Collisionally activated dissociation (CAD) can be effective for top-down LC-MS/MS coupled with high resolution time-of-flight and Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. Moreover, CAD of gas-phase stable noncovalent complexes resulting from electrostatic interactions can be used to probe ligand binding sites (e.g., nucleotides, metals). Electron transfer dissociation (ETD) and electron capture dissociation (ECD) is particularly useful for elucidation of disulfide bonds in large proteins, especially if the electrospray ionization (ESI)-based multiple charging is enhanced (e.g., supercharging). Moreover, ECD and ETD can be used to probe the binding sites of weakly-bound ligands and the topology of protein-protein complexes (e.g., hemoglobin).
Measurement of Trace Components in Biological Fluids using GC-MS: Is it the Way to Go?
Mustafa Ali Mohd (University of Malaya, Kuala Lumpur)
The presentation will focus on the approach of using GCMS for the analysis of complex biological fluids such as blood and urine to measure the presence of trace quantities of contaminants. Some examples that are going to be used are pesticides, Endocrine disrupters such as bisphenol A and some drugs of abuse. The approach of the analytical technique will be elaborated and detailed, as well as the principles involved to maximize the output and sensitivities of the analysis. Some important parameters will be discussed and the comparison with other similar instruments will be made. The discussion will also include the sample preparation steps and the optimization of the analytical technique.
Targeted Mass Spectrometry for the Study of Biological Pathways
Manfred Raida (A-Star Research, Singapore)
In the last years targeted mass spectrometry, or single- or multiple-reaction-monitoring (SRM or MRM), has become a major tool to study cellular systems and to verify biomarker candidates in body fluids. Unlike the usually used Western-Blot approach, MRM allows the selective, quantitative and multiplexed analysis of complex systems. Based on selected proteotypic tryptic peptides, unique peptides, for the proteins to be studied in combination with specific fragments of these peptides generated by collision-induced-fragmentation, the highly specific identification of proteins becomes possible. Cross-reaction as seen with antibodies can be excluded by the selection of proteotypic peptides. This technique requires nano-flow liquid-chromatography coupled to triple-stage-quadrupole mass spectrometers. With internal standards, peptides of identical composition labelled with stable heavy isotopes, an absolute quantification over 3–5 orders of magnitude can be achieved. This allows the analysis of proteins with a low number of copies in a cell. Based on the knowledge of biological pathways, the MRM approach allows the study of actions of drugs on these pathways on the cellular level. In contrast to the usually used antibody-based Western-Blot technique, MRM can be multiplexed to monitor and quantify over 100 proteins or posttranslational modifications in one analytical run. Using protein based prefractionation techniques, as reversed-phase based chromatographic or gel-free electrophoretic techniques, the analysis of low abundant proteins becomes possible. Only the protein-based separation allows the identification of isoforms, splice variants, gene-fusion products, and mutations. The enrichment of specific proteins can be carried out by affinity techniques, based on chemical or immuno-affinities. We will discuss the analysis of cellular systems for proteins in distinct pathways, as the Wnt-pathway, the methods for sample preparation and the approaches for the selection of proteotypic peptides and the corresponding specific fragments.
Perspectives of Qualifications and Validation Activities in a Regulated Laboratory
Shib Mookherjea (Valqual International, USA)
This brief talk is aimed at addressing some of the general elements and tenets of Laboratory Quality Assurance (LQA) while focusing largely on the requirements and implementation of various levels of Qualification & Validation activities and procedures in various Pharmaceutical/Biotech/Medical Devices & Chemical industrial laboratories. This discussion will also encompass the conceptual background of Methods Validation and the Qualification activities for various instruments, systems, and equipment, including HPLC/GC/LC-MS, GC-MS, UV-VIS, etc., from a Risk Assessment and Product Lifecycle perspective. A grassroots, fundamental approach (QbD) to Methods Development & Validation procedures, will be touched upon with reference to Regulatory guidelines (FDA, USP, EP, ICH, WHO, etc). During the discussion the infrastructure of a robust QS compliant and transparent Documentation System with multi-tiered documentation strategies, including Methods Validation Protocols, MVP and SOP requirements at various stages (IQ, OQ, PQ) of deployment for various systems in various R&D, QC, Process Development, Environmental Analysis laboratories for conformance to GLP, GMP , ISO -17025 & other guidelines.
Theory and Practice of Orbitrap Mass Spectrometry
Alexander Makarov (Thermo Fisher Scientific, Bremen, Germany)
The talk oversees short but eventful history of Orbitrap mass spectrometry, from laying down the first principles to its current status in mainstream mass spectrometry. Special emphasis is made on new possibilities opened by recent extension of two major families of instruments, LTQ Orbitrap and Exactive. Future trends and perspectives of the Orbitrap mass spectrometry are discussed, particularly in relation to analysis of proteins.
Applications of IMS TOF in Drug Discovery
Kelly Dong (GSK, China)
The presentation will cover the advantage and challenges of IMS TOF applications in the area of supporting drug discovery. Empowered by the additional dimension of separation, IMS coupling with TOF presents significant technical advancement to assess metabolic soft spot and provide structural guidance for lead optimization process. A good understanding of its challenge and limitation is critical to ensure the quality outcome.
Proteomics Study of Alzheimer's Disease Mechanism: What do Blood Biomarkers Tell Us?
Roman Zubarev (Karolinska Institutet, Stockholm, Sweden)
Alzheimer’s disease (AD) knows no cure, and its origin is still debated. A plausible hypothesis is that the disease is triggered by damaged proteins, but the disease progression may be modulated by the presence or absence of an appropriate phenotype. To test this hypothesis, we developed novel label-free proteomics approaches and investigated blood plasma of 218 individuals in various stages of AD. The main finding is that the healthy controls differ from the individuals with various degrees of dementia by lower levels in blood of proteins damaged by the presence in their sequence of isoaspartic acid (isoAsp). Higher isoAsp levels were found in females than in males, consistent with the prevalence of AD for women. However, no quantitative correlation was found between the isoAsp levels and the AD degree, suggesting that isoAsp in blood could be a risk factor triggering dementia, but not actively participating in its progression. On the other hand, levels of several abundant (‘housekeeping’) proteins could differentiate two early AD stages: stable mild cognitive impairment (MCI) and progressive MCI leading to AD. The total emerging picture supports the hypothesis of dementia being triggered by an elevated risk factor (e.g. IsoAsp) with subsequent disease progression in the presence of a permissive phenotype characterized by certain levels of housekeeping proteins in blood.
The ICH Process, Charter and the Integration with Global Regulatory Organizations
Shib Mookherjea (Valqual International, USA)
Since its inception, The International Conference on Harmonization (ICH), has been recognized as a great resource for International guidance in the process of pharmaceutical development and regulatory implementation. This short presentation will outline the genesis of ICH as an International technical organization, along with the charter purpose and composition of ICH. It will also touch upon the guidance provided by ICH in various aspects of pharmaceutical development including efficacy, stability, purity, potency and overall quality of emerging medicinal products while highlighting the integration of ICH with the current Good Manufacturing Practices (GMP’s) that are prevalent in various ICH tripartite regions.
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