Biophysical Society 67th Annual Meeting Program Guide

In this presentation, we will discuss how the improved IRIS platform can bridge the gap between single-particle detection (’digital’ configu ration) and ensemble reflectance measurements (’analog’ configura tion), creating a new ’hybrid’ system (h-IRIS), providing a substantial improvement in sensitivity, improving the limit of detection by nearly three orders of magnitude. This new hybrid configuration of IRIS combines the advantageous features of high-magnification and low magnifications modalities. Unique capability of the IRIS platform to conduct kinetic binding measurements of individual analytes has been also demonstrated for antibody characterization and a novel competi tion assay for therapeutic antibodies for viral infections. We believe that the scientific community will greatly benefit from our approach of combining SP-IRIS platform and use of whole virus particles for obtain ing competition information for monoclonal antibodies. Speaker Selim Unlu, Distinguished Professor, Boston University 3:30 PM – 5:00 PM HORIBA Scientific A-TEEM Molecular Fingerprinting for Biopharmaceutical Products Such as Vaccines, Insulin, Viral Vectors (AAVs), Cell Media, Exosomes, and More A-TEEM is a novel, robust, and extremely sensitive analytical technique comprising the best attributes of molecular spectroscopy (speed, low per measurement cost, lab-to-line placement) and chromatography (sensitivity, selectivity, low limits of detection and quantification). The A-TEEM technique combines absorbance and fluorescence spec troscopy in one simultaneous measurement, providing five character istics that together create a unique A-TEEM fingerprint: The Extinction Coefficient and Absorbance Spectrum (from absorbance) as well as Quantum Efficiency and the entire Excitation and Emission Spectrum in the form of 3D EEM from fluorescence. The absorbance information is also used to correct the fluorescence EEM for inner filter effects, thus facilitating truly quantitative and highly selectable molecular finger prints. A-TEEM fingerprints of very similar components are often unique, enabling quantitative analysis of responsive components to ppb levels. This makes it a natural fit for the characterization of many relevant bio pharmaceutical products such as Vaccines, Insulin, Viral Vectors (AAVs), Antibody-Drug Conjugates (ADCs), Cell Media, Exosomes, CRISPr/CAS9, and more. As a spectroscopic tool, A-TEEM can be deployed for process analytics, used to monitor bioprocesses, or screen for batch-to-batch variance across a variety of relevant biomolecules. Putting a sample in a cuvette is often the only preparation needed for measurement. For some quantitative measurements, sample dilution may be required. Speaker Linda Kidder Yarlott, Life Sciences Business Development Manager, HORIBA Scientific

the well-based high throughput automated patch clamp system, the SyncroPatch 384. Transporters and ion pumps are transmembrane proteins that facili tate ion flux across biological membranes against their concentration gradient. Solid supported membrane (SSM) electrophysiology using the SURFE2R instruments from Nanion can be used to measure elec trogenic membrane transport, even with low turnover rates, as well as ligand-gated, and leak channels. Any membrane source can be used, meaning that intracellular transporter and channels still present in organellar membrane can be targeted using this method, allowing for direct and functional recordings in a physiologically relevant context. Transporters and intracellular ion channels are increasingly important pharmacological targets for the treatment of a wide variety of diseases. In this presentation, scientist Nataliia Dmitrieva, will present her work on the molecular basis of the function of DgoT, a bacterial homolog of vesicular glutamate transporters. Speakers Andrea Brüggemann, CSO, Nanion Technologies James Costantin, Scientific Market Development Manager – Ion Channels, Eurofins DiscoverX Nataliia Dmitrieva, Forschungszentrum Jülich Jean-Francois Rolland, Head, Electrophysiology, Deputy Director, Discovery Services Axxam SpA 11:30 AM – 1:00 PM axiVEND – iRiS Kinetics Accurate Binding Kinetics for Biological Nanoparticle Analytes Accurate characterization of kinetic interactions between biological ligands and analytes is crucial for research in diagnostics and therapeu tics. Application areas of label-free kinetics are broad – ranging from drug and antibody screening to characterization of viral carriers for gene therapy and vaccine development. For these diverse applications, the size of biological analytes has a size range spanning many orders of magnitude. Label-free sensing platforms and sensor chips are often optimized for the type of analyte target, for example, Surface Plasmon Resonance (SPR) sensors have thickness and density of the surface hydrogel chemistry optimized for detection of analytes based on their molecular weights. Here, we present a sensor platform that provides accurate binding characterization of broad range of analytes and com bines label-free detection of ensembles and single particles on a single reader instrument. Interferometric reflectance imaging sensor (IRIS) technology is based on interference of light from an optically transparent thin film—the same phenomenon that gives rainbow colors to a soap film when illu minated by white light. IRIS has two modalities: (i) low-magnification (ensemble biomolecular mass measurements) allowing for multiplexed affinity measurements and (ii) high-magnification (digital detection of individual nanoparticles). Single particle IRIS has been demonstrated for multiplexed detection and characterization of individual biological nanoparticles (BNP) such as extra-cellular vesicles and virions in com plex samples using disposable cartridges. Low magnification IRIS offers large field of view and ability to simultaneously quantify binding of ana lytes to highly multiplexed probe molecules arrayed on the sensor sur face. However, accurate characterization of large analytes (50-150nm size) presents challenges due to non-linearity of the optical interfer ence signal with surface binding of sparse BNP analytes. We have recently demonstrated that the affinity of large analytes (extracellular vesicles) can be characterized, qualitatively, on IRIS chips benefiting from IRIS’ capability of uniform illumination at different wavelengths.

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