|Building a digital pathology ecosystem for education and research|
Yves Sucaet, Silke Smeets, Stijn Piessens, Sabrina D'Haese, Chris Groven, Wim Waelput, Peter In't Veld
We wanted to build a core digital pathology infrastructure to support different use cases. Various images platforms needed to be accessible through a single access point, and support different user profiles. We wanted a scalable solution that would allow interaction between equipment from different research groups.
We built a centralized infrastructure that integrates a variety of imaging platforms, and now have an interconnected network of heterogeneous and scalable information silos.
|Sample Stabilization and Purification Devices for Superior Forensic DNA Storage, Purification and Profiling|
Geoffrey Routh,& Bassam El-Fahmawi
The tools used for evidentiary sample collection are often challenged in criminal cases. Maintaining the integrity of evidentiary samples continues to be a major challenge - especially when samples stay in transit during transportation where sample integrity is highly compromised. Thus, there is a crucial need for a cost-effective, efficient DNA collection and storage system that maintain sample integrity from collection until processing, while providing a means to ensure proper chain of custody
|A Simple Plate Based Assay Using pH Sensor Dye to Screen for Internalizing Antibody|
Nidhi Nath, Becky Godat, Cesear Corona, Chad Zimprich, Mark McDougall, Poncho Meisenheimer, Marjeta Urh
Receptor mediated internalization is a key mechanism of action (MOA) for antibody drug conjugates (ADCs). However, current methods of studying antibody internalization have several limitations including: 1) A multistep process not suitable for screening; 2) Low signal-to-background ratios; 3) Not suitable for kinetic measurements. We have developed a method that mitigates problems associated with traditional internalization assays.
|An IgG Cleaving Protease from S equi with Improved Activity Against Mouse IgGs|
Chris Hosfield, Philip Compton, Luca Fornelli, Paul Thomas, Neil L. Kelleher, Michael Rosenblatt & Marjeta Urh
Here we have expressed and purified a modified recombinant IdeZ, and show that it has significantly improved activity against mouse IgG2a and IgG3 subclasses when compared to IdeS. We also demonstrate the use of IdeZ in LC-MS workflows for human and mouse IgG characterization.
|Minimizing Carry-over for High Throughput Analysis|
Christian Berchtold1, Reto Bolliger2, Guenter Boehm2, Götz Schlotterbeck1
Minimal carry-over is a prerequisite for high throughput analysis. However, minimized carry-over and cycle time are competing and a careful optimization is mandatory. In this study the influence of wash conditions on carry-over of various compounds was investigated. A strategy to minimize carry-over was developed. The influences of different wash tasks were investigated. Finally the contribution of different system components such as injector valve or column was studied.
|A Comparison of ITEX Dynamic Headspace–GC/MS to other Enrichment Techniques for Analysis of Flavoring Compounds|
Douglas Doster1; Roger Pearson1; Sean Eppel1; Ken Rice1; Tom Flug2; Brian Peat2; Guenter Boehm2
Enrichment techniques are commonly used for the analysis of flavoring compounds in different matrices with GC/MS. Analysis of flavoring compounds is done by purge & trap, SPME or headspace, depending on requirements for sensitivity. The In-Tube Ex¬traction (ITEX) Dynamic Headspace uses a micro trap filled with an adsorbent material to efficiently extract the compounds. Here we evaluate if the ITEX can be used to effectively analyze for these compounds and reduce the analyst’s time involved.
|Automated in-gel digestion on a commercial autosampler directly coupled to nanoLC-MS/MS|
Achermann François, Bolliger Reto, Buchs Natasha, Doiron Nicholas, Lagache Braga Sophie, Heller Manfred, Boehm Guenter
SDS-PAGE separates protein samples from LC-MS incompatible contaminations, and is frequently used to fractionate proteins of entire proteomes. One disadvantage is that gel lanes have to be cut into many slices, followed by in-gel digestion of proteins and extraction of peptides. The number of these gel slices goes into the hundreds, rendering this process very repetitive and prone to mistakes and errors during sample handling. Automation reduces such risks and improves reproducibility.
|Automated sample preparation workflows for quantitative proteomics applications|
Oliver Popp1, Lucas Luethy2, Tamara Kanashova1, HaAn Nguyen1, Julia Kikuchi1, Guenter Boehm2, Thomas Blenkers3, Andreas Bruchmann3, Gunnar Dittmar1
Mass spectrometry based proteomics requires large scale identification of peptides, and depends upon efficient sample preparation. Recently, we presented two automated protein-digestion setups, in-solution and in-gel digestion. We extended these techniques by implementing dimethyl labelling (DML). Furthermore, we established an automated phospho-peptide (PP) enrichment procedure in a 96-well formate, generating phospho-proteomic data in very short time.
|Optimization of a Vacuum Ultraviolet Photoionization source for Gas Chromatography used with a High Resolution Time of Flight Mass Spectrometer|
Lloyd Allen and Viatcheslav Artaev
-Tune solution allows optimization of ion source parameters for
both proton transfer and direct ionization
-Independent ionization processes exist for M+ and MH+
-Optimizing for dopant signal intensity yields inferior results
-Degree of fragmentation remains relatively constant over a
range of source conditions