3rd NovAliX Conference

Biophysics in Drug Discovery 2016

Developing the Synergy between Biophysics and Medicinal Chemistry to Deliver Better Drugs

 Strasbourg, France    June 7-10, 2016


Real-time, Solution-based Measurement of Protein Conformational Change

Structural information is critical to understand the function or dysfunction of a protein and the mechanism of action of a drug targeted to that protein.  Existing tools for measuring protein structure are expensive, slow, and labor intensive.  Drug discovery and biological research have a pressing need for improved protein analysis tools with greater sensitivity, lower cost, and high throughput. The Biodesy Delta is a revolutionary system that uniquely enables real-time measurement of protein conformational change by binding an analyte. The Delta can work with any protein, regardless of size and any ligand, including small molecules, fragments, proteins and mixtures.


A Visual Approximation to On-the-Fly Computational Thermodynamic Profiling

ΔG = -RT lnK; if there were a "visualizable" approximation to K, we could indeed help drug designers steer their affinities towards a desired thermodynamic pro-file, e.g. more entropic contribution. In a collaboration with Bayer and Hamburg University we have written a novel software that helps understanding the ener-getic consequences of balancing H-bonds versus desolvation penalties.
The system is embedded in SeeSAR, and it is programmed such that the effects can be visualized, semi-quantified, and rationalized at an atomic level. Additional visualization includes a statistics-based torsional alerting system, and an optimization of the H-bond network in milliseconds. We will briefly outline the theory behind and give several examples.

Bruker BioSpin

Fragment Screening by Ligand Observed NMR

Ligand observed NMR is a powerful technique for the screening of fragment like small molecules to biomolecular targets in solution. In recent years methodological and technical advancements have enabled NMR based fragment screening to be performed in full automation and with significantly reduced consumption of unlabeled target protein and fragments. Here we present practical aspects on how to setup NMR based ligand observed screening experiments in an automated fashion for the three basic experiments: STD, wLOGSY and relaxation based methods. Furthermore, we show optimized measurement parameters, tips for sample preparation, tool samples to setup and test NMR screening and, lastly, miscellaneous tools, scripts etc. for automation. The goal is to enable the drug discovery community to setup those experiments and assays on available instrumentation and to reduce the hurdle for non-experts to use NMR screening for Fragment Based Lead Discovery (FBLD).


The WAVE of the Future in Kinetics

By combining a level of sensitivity superior to Surface Plasmon Resonance (SPR) with a crude sample robustness normally only achieved with plate-based assays, the Creoptix™ WAVE system is revolutionizing the study of molecular interactions and changing the world of drug discovery.
Engineered around a proprietary waveguide interferometry technology, the WAVE system offers superior data quality across the broadest range of compounds for unrivaled flexibility and unsurpassed insight—all made possible by the instrument’s three core attributes: enhanced sensitivity, fast kinetics (off-rates of 5/sec) and a disposable and most robust microfluidics compatible with crude samples.
So welcome to the future kinetics analysis. Where will the WAVE take you?

Dynamic Biosensors

Biophysical Analysis of Molecular Interactions and Enzymatic Activity with switchSENSE

switchSENSE® is an automated biosensor chip technology that employs electrically actuated DNA nanolevers for the real-time measurement of binding kinetics, affinities, and enzymatic activities. Interactions between proteins, DNA/RNA, and small molecules can be detected with femto-molar sensitivity. In the same workflow, protein diameters are analyzed with Angstrom accuracy enabling the measurement of even minor conformational changes.
In this talk we will present novel applications of switchSENSE including the characterization of the CRISPR/Cas9 system for genome editing, and the detection of conformational changes in proteins that are elicited by small molecules.

Malvern Instruments

Innovative Solutions for Biosimilars

Once a potential therapeutic biologic has been Identified, the process of developing the therapeutic continues through costly, complex and increasingly demanding phases. Candidates and formulations that fail to satisfy the predetermined criteria are removed from the main product pipeline and placed in the support pipeline area. More in-depth analytical data is needed to understand the reason for such failures. Based upon this data, failure predictive analytical assays are developed to ensure the most relevant tests are introduced to decrease drug development cost. Such innovative analytical approaches are also of great interest in the growing Biosimilar market where Biosimilar/Innovator products undergo side by side extensive testing.

In this context, Malvern has established in 2012’s a Biosciences Development Initiative (BDI), partnering with industry and academia to rapidly identify and assess analytical problems and deliver innovative solutions. Several of these technologies related to particle size, viscosity and structural integrity will be presented and mainly:

  • The Viscosizer TD: Small volume both viscosity and size screening method compatible with complex media using Taylor dispersion.
  • The HELIX a hyphenated DLS-Raman system that provides insight into stability at the microscopic structural level and develops mechanistic understanding of protein aggregation.

Molecular Sensing

Affinity Assays for GPCR ligand binding using TruBind™ Molecular Interaction Analysis

Integral membrane proteins such as G-protein coupled receptors (GPCRs) are relevant targets in drug discovery. However, biophysical investigation of isolated and purified membrane proteins is challenging because of low expression levels in commonly used expression systems and instability of the proteins in a non-native environment. Therefore, developing techniques to study integral membrane proteins in their native and complex membrane environment is of high importance.
Based on the principle of Back-Scattering Interferometry, TruBind Molecular Interaction Analysis is a label-free technique for mass- and matrix independent biophysical characterization of small molecule interaction with complex integral membrane proteins in a native-like environment. TruBind analysis enables direct determination of compound affinity without the use of labels or tethering the target to a surface.

Using TruBind analysis, we have investigated the binding ability of antagonistic ligands to a thermostabilized and truncated version of a GPCR in a detergent purified and in a membrane bound state. In addition, we also tested the binding of the same ligands to the corresponding wild type GPCR in a membrane bound state.

NanoTemper Technologies

Prometheus NT.48 - The Stability Expert

The Prometheus NT.48 with nanoDSF technology is the ideal instrument for rapid and precise thermal stability screening in biopharmaceutical development. The innovative features of this instrument allow for markedly fast analysis times and lower sample volumes. In a new study in collaboration with Coriolis Pharma, we demonstrate comparable precision and consistency in data acquired from both nanoDSF and the "gold standard" µDSC methodologies.
In addition to speed, precision and throughput, nanoDSF is a robust method that does not require laborious sample preparation such as dialysis or filtration. It works in any buffer, even with detergents, and tolerates high viscosity samples that are typical in formulation studies.
Performing nanoDSF with the Prometheus NT.48 helps users overcome some of the cumbersome drawbacks associated with µDSC: nanoDSF provides perfect ease of use, is 100 times faster and requires 40 times less sample!
The self-filling capillaries on the Prometheus NT.48 allow for easy sample handling, even for highly concentrated and very viscous samples. Additionally, the maintenance-free instrumentation does not require laborious equilibration and washing.


Balancing Standard and Unique Services for Drug Discovery

An overview of NovAliX' chemistry and biophysical services with particular focus on the unusual or unique platforms including native MS; chemical micro-arrays and electron microscopy.

Sierra Sensors

Introducing MASS-2 for High Performance, High Throughput SPR Analysis

High-throughput, high-performance SPR analysis of compound and fragment binding interactions using the new MASS-2 analytical biosensor from Sierra Sensors GmbH will be presented.  The MASS-2 is a 32-sensor, 8-channel, continuous flow biosensor.  Capable of processing 8 samples simultaneously, the MASS-2 can screen 3300+ sample solutions per day, generating over 10400 control-subtracted interactions.  Able to robustly measure the binding of fragments with a mass as small as 100 Da, the MASS-2 is also ideal for analysis of large proteins and crude samples.  Individual needle control means the MASS-2 can be a 1- to 8-channel biosensor on command, and the 4-buffer continuous flow pump means completely different assays or assay conditions can be run simultaneously.  Examples of screening and kinetic characterization of low-molecular-mass compound binding interactions using MASS-2 will be reviewed.