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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.

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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.

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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.

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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.

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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.

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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.

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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

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High Performance Comprehensive Two-Dimensional Gas Chromatography Coupled with a High Resolution Multi-Reflecting TOFMS for Confident Non-Target Analyte Identification
Scott J. Pugh, Viatcheslav Artaev, Mark F. Merrick, Jack Cochran

The use of comprehensive two-dimensional gas chromatography to help increase chromatographic resolution is a major step in tackling the problem of confident peak identification in a complex sample matrix. Combining the separation power of two-dimensional gas chromatography, with resolving power greater than 25,000, and sub ppm mass accuracies of a high resolution multi-reflecting TOFMS is the ideal solution to confident compound identification within a complex sample matrix.

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Fragmentation Trees for Automated de novo Interpretation of Impure Electron Ionization Spectra from Gas Chromatographic Complex Mixture Analysis—Chemical Deconvolution
Kevin Siek, Vasily Makarov, Viatcheslav Artaev, Dmitry Mazur, Albert T. Lebedev

-Chemical deconvolution algorithms accurately reported independent
components of dead coelutions where such components belong to distinct chemical classes.
-Present algorithms fail to distinguish chemically similar analytes such as branched and linear alkanes, thus chemical deconvolution is not a
substitute for GCxGC.
-Chemical deconvolution algorithms accurately rejected more than 80% of spurious signals from manually curated spectra found to be contaminated with unrelated signa

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Effective Comparison of Yeast Extracts Using High Resolution GC and GCxGC-HRTOFMS
David E. Alonso and Joe Binkley

The Pegasus GC-HRT and GC-HRT 4D instruments are effective tools for the comprehensive analysis of yeast extracts.
• Enhanced chromatographic resolution of the HRT 4D system facilitated compound characterization by improving spectral similarity scores
• High quality data resulted in excellent spectral similarity scores when compared to large, well-established databases (NIST, Wiley)
• High resolution, accurate mass data was crucial for determination of fragment, molecular, and adduct

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Scientific News
Portable Kit Can Recover Traces of Chemical Evidence
A chemist at the National Institute of Standards and Technology (NIST) has developed a portable version of his method for recovering trace chemicals such as environmental pollutants and forensic evidence including secret graves and arson fire debris.
Forensics Professor Detects Blood on Revolutionary War Projectiles
More than 230 years after the Revolutionary War ended, Edinboro University professor of forensic science Dr. Ted Yeshion has found the presence of blood on buckshot recovered from a battlefield in upstate New York.
Single Molecule Detection of Contaminants, Explosives or Diseases
A technique that combines the ultrasensitivity of surface-enhanced Raman scattering (SERS) with a slippery surface invented by Penn State researchers will make it feasible to detect single molecules of a number of chemical and biological species from gaseous, liquid or solid samples.
Potential New Tool for Forensic Science
Microbial communities associated with humans tick in predictable, clock-like succession following death.
Perfecting Age Estimations Under 25
The Idaho State University Department of Anthropology has received a $510,409 grant from the National Institute of Justice to develop forensic science techniques to better identify individuals under 25 years of age for criminal justice purposes.
Identifying Gender from a Fingerprint
Culprits beware, a University at Albany research group, led by assistant chemistry professor Jan Halámek, is taking crime scene fingerprint identification to a new level.
Viruses, Too, Are Our Fingerprint
A group of researchers from the University of Helsinki and the University of Edinburgh have been the first to find the genetic material of a human virus from old human bones.
Questioning the Validity of Forensic DNA Match Statistic
Fifteen years of criminal cases with affected mixture evidence.
Study Raises Questions About DNA Evidence
University of Indianapolis researchers say contamination through secondary transfer of material could implicate the innocent or help the guilty go free.
'Forensic Toolkit’ to Improve Evidence Detection and Analysis
Students from The University of Dundee have been developing a forensic `toolkit’ that will allow investigators to determine the age of fingerprints, detect traces of steel on bone from stabbings, and produce a biosensitive spray that can reveal traces of bodily fluids at crime scenes.
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