SLAS2022

On-demand

Introduction

Introduction - SLAS2022

If you did not make it to Boston or could not visit us during SLAS2022 to learn about our latest Bruker’s Surface Plasmon Resonance (SPR) instruments, Mass Spectrometry solutions for high throughput screening you still have time to listen to some of our presentations during the Conference.

Listen On-Demand to Cyrill Brunner, who presented his latest work titled “Information-Rich High Throughput Screening with Multiplexing Surface Plasmon Resonance - using the Bruker SPR-32 Pro. His talk was held during the scientific session “Biochemical and Biophysical Assays on February 7, 2022.  

Also On-Demand are several presentations from Tuesday’s Exhibitor Tutorial.  We hope you find the presentations informative and look forward to hearing from you if you have any questions.  See you in 2023!

Scientific Presentation: February 7, 2022

Scientific Presentation: Monday, February 7, 2022
5:00 pm – 5:30 pm (EST)

Title: Information-rich High Throughput Screening with Multiplexing Surface Plasmon Resonance
Speaker: Cyrill Brunner, Ph.D.

Abstract:
Surface Plasmon Resonance is an established and widely used biophysical technology in screening and lead development campaigns for novel drug candidates. The real-time, label-free analysis of interactions offers additional insights into kinetics unlike endpoint assays like fluorescence-based assays. Multiplexing SPR systems further allow to study the interaction of a drug candidate against multiple targets simultaneously; a task which requires multiple repetitions of the same screening with other techniques. However, the throughput is often comparably low and the number of targets in a fast, efficient quantitative screening campaign is severely limited.

We present an assay set-up that allows the binding assessment of a single analyte against dozens of targets or the affinity determination of one analyte against multiple targets with a single injection. Partnered with our established SPR detection system, it’s possible to perform measurements also at high molecular weight differences between analyte and target of 10-3. Furthermore, this assay set-up facilitates the determination of thermodynamic constants at low running times due to parallel readout. Access to this information is typically relevant in subsequent lead development processes.

We tested the assay set-up with multiple established assay modes for two model cases. The interaction of oligonucleotides against complementary strands and of a set of sulphonamides against multiple carbonic anhydrase isozymes confirmed the performance and robustness. Additionally, the sensitivity of our novel assay set-up was assessed with an interaction of DNP-modified amino acids (241-283 Da) against specific antibodies (150 kDa).

We herewith present a SPR assay set-up with industry-leading throughput in affinity determination and multiplexing capabilities for all types of analytes. Thus, the extension of typical parameters from a SPR experiment by a quantitative selectivity assessment allows for a more informed hit selection process in screening campaigns.

Cyrill Brunner, Ph.D.
Application Specialist, Bruker Switzerland, Fällanden, Switzerland

Dr. Cyrill Brunner is a trained pharmacist and obtained his Ph.D. at ETH Zurich in the group of Prof. Gisbert Schneider. His main background are numerous biophysical assays including SPR and computational drug design. He joined Bruker as application specialist in April 2020 and heads the SPR application lab in Fällanden, Switzerland.

Bruker Exhibitor Tutorial: February 8, 2022

Bruker Exhibitor Tutorial: Tuesday, February 8, 2022
2:30 pm – 4:00 pm (EST)

Agenda

2:30 – 2:35   Michael Greig, Ph.D.: Introduction

2:35 – 2:50   Prof. Matthias Trost: MALDI-TOF assay identifies nilotinib as an inhibitor of inflammation in acute myeloid leukemia

2:50 – 2:55    Q&A

2:55 – 3:10    Arndt Asperger, Ph.D.: 
timsTOF MALDI PharmaPulse: A next-generation MS based label-free uHTS solution taking advantage of timsTOF technology


3:10 – 3:15    Q&A

3:15 -  3:30    Sven Malik: Next level of high throughput SPR screening: 10,000 compound screen enabled by Bruker SPR

 

3:30 – 3:35    Q&A

3:35– 3:50     Prof. Philip D. Compton: A Gateway to Mass Spectrometry: The SampleStream Platform for Biomolecular Purification

3:50 – 4:00    Q&A

 

Tutorial Description: Introducing Unbiased, Deep HTS by label-free timsTOF Mass Spectrometry (MS) and Surface Plasmon Resonance (SPR)
Speaker: Meike Hamester, Ph.D., Michael Greig, Ph.D.

Introducing for the first time the capability of deep data coverage for HTS and uHTS by the all new timsTOF MALDI PharmaPulse Mass Spectrometer (timsTOF MPP). At SLAS Bruker is launching a new instrument platform combining HTS and uHTS capabilities of MALDI Pharma Pulse with deep data mass spectrometry by high-resolution QTOF and trapped ion mobility separation (TIMS). 

Furthermore, Bruker is presenting it's Surface Plasmon Resonance (SPR) platform, the SPR Pro Series delivering unprecedented data quality and throughput. 

Learn in our Bruker Exhibitor Tutorial how label-free techniques such as mass spectrometry and surface plasmon resonance (SPR) can enhance your high-throughput screening results without compromising on cost or throughput.


Title:  MALDI-TOF assay identifies nilotinib as an inhibitor of inflammation in acute myeloid leukemia
Speaker: Prof. Matthias Trost, Ph.D.

Abstract:
Inflammatory responses are important in cancer, particularly in the context of monocyte-rich aggressive myeloid neoplasm. We developed a label-free cellular phenotypic drug discovery assay to identify anti-inflammatory drugs in human monocytes derived from acute myeloid leukemia (AML), by tracking several biological features ionizing from only 2,500 cells using matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) HTS mass spectrometry (Bruker rapifleX MPP).

A proof-of-concept screen showed that the BCR-ABL inhibitor nilotinib, but not the structurally similar imatinib, blocks inflammatory responses. In order to identify the cellular (off-)targets of nilotinib, we performed thermal proteome profiling (TPP). Unlike imatinib, nilotinib and other later generation BCR-ABL inhibitors inhibit the p38α-MK2/3 signalling axis which suppressed the expression of inflammatory cytokines, cell adhesion and innate immunity markers in activated human monocytes derived from AML. Thus, our study provides a tool for the discovery of new anti-inflammatory drugs, which could contribute to the treatment of inflammation in myeloid neoplasms and other diseases.


Title: timsTOF MALDI PharmaPulse: A next-generation MS based label-free uHTS solution taking advantage of timsTOF technology
Speaker: Arndt Asperger, Ph.D.

Abstract:
Mass spectrometry (MS) offers huge potential for enhancing the efficiency of early drug discovery.

In this workshop, we will introduce the timsTOF MALDI PharmaPulse (MPP), a next-generation label-free solution for MS based HTS and uHTS taking advantage of Bruker´s innovative timsTOF technology.
timsTOF MPP combines proven speed and robustness of MALDI with trapped ion mobility (TIMS) separation and high-resolution, accurate-mass time-of-flight (TOF) MS detection, allowing for new, deeper insights into molecular interactions at uHTS compatible reading rates.

Advanced timsTOF MPP operation modes provide enhanced assay specificity and thus, open up exciting new HTS possibilities. Analysis in MS/MS mode utilizes compound-specific fragment ions resulting in expanded range of quantitation. TIMS offers an additional dimension of gas-phase separation for efficient reduction of isobaric or isomeric background interferences running significantly faster than any LC/SPE based separation method currently available.

Key features described above, make the timsTOF MPP a highly versatile platform for a broad range of HTS and uHTS applications in early drug discovery, including biochemical and cell-based mechanistic assays, cell-uptake assays and binding assays. In addition, timsTOF MPP serves as a highly capable platform for near real-time verification of synthesis products obtained from high-throughput experimentation (HTE) chemistry in search of new drug molecules.

timsTOF MPP solutions includes dedicated MALDI PharmaPulse HTS software, guiding the user through setup and execution of various screening workflows, featuring an automation interface for seamless integration with lab automation systems and providing a customizable export interface for transfer of data and results to external downstream analysis software.


Title:  Next level of high throughput SPR screening: 10,000 compound Screen enabled by Bruker SPR
Speaker: Sven Malik

Abstract:
Even as researchers leverage a host of new technologies to pervade the design of libraries and screening workflows for a detailed candidate selection, it has remained a challenge to generate real-time, direct-binding results at scale despite the confidence and insights that they provide.

The Bruker SPR-32 Pro platform was designed to empower researchers with the sensitivity and multiplexing capabilities needed to support and expedite these evolving discovery cycles for both fragments and biologics. This talk highlights the key points to enable fragment screens at a new throughput levels. An entire library of 10,000 compounds (28x384-well plates) was comprehensively screened for general-binding over 5-6 days against three targets and a reference per channel. Plate loading was fully automated using the Orbitor plate changer (Thermo Scientific).


Title: A Gateway to Mass Spectrometry: The SampleStream Platform for Biomolecular Purification
Speaker: Prof. Philip D. Compton, Ph.D.

Abstract:
Antibody- and other protein-based therapeutics require extensive and varied characterization throughout discovery and development. Traditionally, various chromatographic methods combined with mass spectrometry have served well for many of the required analytical targets. However, some limitations of chromatography either create upstream sample processing challenges or render certain workflows impossible.

The first major challenge is the limited loading volume for all chromatographic types. While isocratic elution in reversed-phase imposes a limit on loading volume, the challenge is most keenly felt in size exclusion applications where loading volume is limited to a fraction of the total column volume. The second major challenge is throughput. Due to column re-equilibration, LC cycle times are generally limited to a minimum of 4-5 min/sample. Combined, these limitations force the addition of pre-concentration steps and limit throughput.

The SampleStream Platform for Biomolecular Purification provides a unique alternative that directly addresses these challenges. Functioning essentially as an online molecular weight cutoff spin filter, SampleStream’s fluidic channel can concentrate arbitrarily large volumes of arbitrarily dilute solution, all while delivering efficient buffer exchange into either denaturing or native buffers. The flow rates utilized for concentration and buffer exchange are much higher than typical LC flow rates, while the elution flow rate matches commonly used 2.1 mm chromatographic methods. This enables the SampleStream Platform to deliver cycle times as fast as 30 sec/sample.

Here, we will present data from the SampleStream Platform coupled with a Bruker timsTOF Pro 2 on representative mAb and ADC molecules to demonstrate the exceptional data quality that can be achieved with this hardware platform.

Meike Hamester, Ph.D.
Director label-free technologies, Bruker Daltonics GmbH & Co. KG, Bremen, Germany

 

Michael Greig, Ph.D.
Director Pharma/Biopharma Americas, Bruker Daltonics, Billerica, MA, USA

Prof. Matthias Trost, Ph.D.
Newcastle University, Faculty of Medical Sciences, Newcastle, UK

Matthias Trost is a proteomics expert with over 20 years of experience in mass spectrometry. He studied chemistry in Freiburg, Germany, and Manchester, UK. After his Ph.D. in Cellular Microbiology and Proteomics at the Helmholtz Centre for Infection Biology in Braunschweig, Germany, he spent five years as a postdoc at the Institute for Research in Immunology and Cancer in Montreal, Canada. In 2010, he became PI and Head of Proteomics at the MRC Protein Phosphorylation and Ubiquitylation Unit at the University of Dundee. In 2017, he was appointed Professor of Proteomics at Newcastle University.

Since 2019, he is a Wellcome Trust Investigator. Matthias has a substantial expertise in the proteomics analysis of post-translational modifications, particularly phosphorylation and ubiquitylation. His main biological interest is in innate immunity signalling and macrophage biology. In recent years, his lab has increasingly focused on using mass spectrometry for drug discovery. For this, they pioneered the usage of high-throughput MALDI-TOF mass spectrometry for drug screening. Recently, he was awarded the Biochemical Society’s 2022 Industry and Academic Collaboration Award.

Arndt Asperger, Ph.D.
Senior Applications Scientist, Bruker Daltonics GmbH & Co. KG, Bremen, Germany

Arndt Asperger, Ph.D., is working as a senior applications scientist at Bruker Daltonics GmbH & Co. KG, Bremen, Germany. Arndt is analytical chemist by training and got his Ph.D. from Leipzig University. He joined Bruker in 2002 and has been working in various positions related to MALDI-MS applications and applications development. His current focus is on applications development of MALDI based high-throughput MS methods.

Sven Malik
Senior Application Specialist, Bruker Daltonics SPR, Hamburg, Germany

Sven studied Bioprocess Engineering at the Technical University of Dresden, Germany and graduated in 2012. During his diploma studies, he developed inverse assays for capture-molecules using surface plasmon resonance. Thereby, he focused on aptamers to establish a monitorable SELEX-process. Additionally, he investigated the interaction of cytomegalovirus protein pp65 against certain antibodies.

In 2012, he joined Sierra Sensors GmbH to start his role as application specialist. After 6 years he became senior application specialist as Bruker acquired Sierra Sensors GmbH. Currently he is involved in the customer support and product development.

Prof. Philip D. Compton, Ph.D.
Research Associate Professor, Department of Chemistry, Northwestern University; CEO, Integrated Protein Technologies, Inc., Evanston, IL, USA

Philip D. Compton received his Ph.D. from the University of Virginia while working under Prof. Donald F. Hunt.  His graduate work culminated in the invention and implementation of a discharge-based reagent ion source capable of generating reagents for electron transfer dissociation and proton transfer reactions as well as the discovery of a new ETD reagent with favorable properties. The source has now been commercialized by Thermo Fisher Scientific as the Easy-ETD reagent ion source, implemented on their Tribrid series of mass spectrometers. 

After his time in Virginia, Phil moved back home to Chicago, to join Prof. Neil Kelleher at Northwestern University.  For more than a decade as a research assistant professor, Dr. Compton has maintained Northwestern Protoemics’ research infrastructure, trained new users, and lead the development of new technologies. Dr. Compton has since transitioned to CEO of Integrated Protein Technologies where he leads the development of the SampleStream platform for biomolecular purification. Currently, he is working to develop workflows of interest that take advantage of SampleStream’s unique capabilities and to officially launch the platform with OEM channel partners.

Spotlight Session: February 8, 2022

Spotlight Sessions

Tuesday, February 8 – 1:00 – 1:20 pm (EST)

Title: Bead Assisted Mass Spectrometry Platform with High-Throughput MALDI MS Detection for Targeted Assays

Speaker: Sergei Dikler, Bruker Scientific, LLC, Billerica, MA, USA

Bead Assisted Mass Spectrometry (BAMS™) is an assay platform that combines immuno-affinity capture on magnetic beads with high-throughput, label-free detection by MALDI-TOF mass spectrometry for multiplexed, targeted proteomic assays. Almost any protein target is accessible as long as a specific affinity reagent (i.e., antibody) for it is available.

This presentation will focus on the detailed description of the workflow and examples of BAMS™ assays developed for targets relevant to neurology, epigenetics, and food allergen testing. 

Sergei Dikler, Ph.D.
Technical Project Manager, Pharma Business Area
Bruker Scientific, LLC, Billerica, MA, USA

文学

 

For Research Use Only. Not for use in clinical diagnostic procedures.