Digital SPR™ is a next-generation technology for characterizing biomolecular interactions. By combining digital microfluidics (DMF) with SPR, digital SPR systems can effectively bypass common challenges found in traditional SPR and BLI platforms.
Understanding drug mechanisms in complex systems, particularly in the brain, remains challenging despite advancements at the molecular level. Yet, cell-specific technologies like DART provide a promising solution, enabling targeted drug delivery to genetically defined neuronal cell types, thereby allowing the observation of the drug effects on brain dynamics and animal behavior. (Shields et al., bioRxiv, 2022, Shields et al., Science, 2017).
The underlying principle of DART technology is the covalent interaction between a protein and its target chemical molecule. In this webinar, we will discuss the optimization of the covalent capture affinity of DART, and the subsequent functional analysis of binding kinetics, using digital SPR.
– What digital SPR is and how it works.
– See how data quality and throughput are maximized using the latest advances in digital SPR to eliminate artifacts, optimize accuracy, and automate tedious, error-prone sample preps.
-See how researchers at Duke University utilize this technology to rapidly assess the binding kinetics of a covalent interaction between a biological ligand and its chemical analyte.
As the world’s only surface plasmon resonance (SPR) system powered by digital microfluidics, Alto™ revolutionizes real-time interaction analysis by eliminating the need to compromise on quality and time. Go from sample to answer within hours while streamlining even the toughest of biologics applications with Alto’s intuitive and automated ecosystem. Designed to take the complexity out of SPR, Alto™ will empower your team with the data they need to take their discoveries to the next level.
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Obtain high-quality data and publish with confidence using OpenSPR®, the world’s first benchtop surface plasmon resonance (SPR) instrument. For a fraction of the cost of existing solutions, OpenSPR® provides real-time, label-free analysis of affinity and kinetics data for a wide range of biomolecular applications. Our unique nanotechnology-based sensors, which produce a localized SPR (LSPR) phenomenon, work seamlessly with an intuitive software interface and robust hardware to generate publication-quality data. Choose between OpenSPR®, a two-channel solution ideal for new or occasional users, and the OpenSPR-XT™, an automated system for users looking to upgrade to hands-free operation.
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Oligonucleotides offer a promising vector for the treatment of diseases related to gene expression and have recently transformed the vaccine industry with the resounding success of mRNA-based COVID-19 vaccines. In this application note, we demonstrate Alto’s applicability to the development of oligonucleotide systems in various therapeutic capacities. Alto is used to accurately and precisely measure affinities and provide on/off-rates equivalent to those obtained with conventional SPR.
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Many human diseases are known to be associated with transmembrane proteins (TPs), making them ideal candidates for drug development. Multi-pass transmembrane proteins are not stable outside of the cell membrane environment, and are therefore difficult to purify and to express in large quantities. This has resulted in the under-characterization of a critical group of biomolecules. One strategy for overcoming these challenges is to use vectors that closely mimic cell membranes, such as nanodiscs. In this application note, Altoâ„¢ Digital SPR was used to characterize binding kinetics of Claudin-18.2 in nanodiscs from ACROBiosystems, demonstrating its ability to provide high-quality data while significantly reducing precious sample consumption and time to results.
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Screening is a common tool used to quickly test activity of a large number of chemical or biological compounds. Commonly seen as one of the first steps within the drug discovery process, screening assays are used on large scale libraries to quickly test and select candidates that bind to or affect a desired target. Surface plasmon resonance (SPR) is often used to conduct high-throughput screening to test for real-time binding activity of a variety of biological and chemical compounds.
Direct screening is one of the high-throughput assays offered on Alto, Nicoya’s digital microfluidics (DMF) based SPR instrument. Alto’s screening protocol allows users to establish binding activity of up to 48 unique analytes against desired targets in less than 4 hours. This technical note will go through two separate studies, with one demonstrating yes/no binding of various ligands and analytes, and the other presenting how Alto’s screening protocol can be used for the discovery of new therapeutic antibodies. The below case studies exemplifies five main qualities of Alto’s screening protocol:
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Rapid changes in influenza antiviral target proteins due to antigenic drift result in cloaking of the influenza virus from the immune system of vaccinated hosts. Hence, annual formulation updates for influenza vaccines and related antibody therapies are required to preserve immune recognition against different influenza subtypes. As such, characterizing the binding kinetics and epitope diversity of various antibodies to influenza viral antigens is essential for treating and preventing potential outbreaks. In collaboration with Sino Biological, a global leader in recombinant technology, we use the Nicoya Alto® digital surface plasmon resonance (SPR) system to perform kinetic analysis and epitope characterization of 16 antibodies against Influenza A hemagglutinin (HA), using only 1 µg of antigen and 100 ng of antibody. Altoâ„¢ now is introduced with a simplified 16×16 epitope binning application for biotherapeutics research, galvanizing your productivity by reducing your cost and effort.
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As the first digital microfluidic (DMF) powered surface plasmon resonance (SPR) instrument in the market, Alto™ provides users with a quantitation assay to quantify proteins of interest in various mediums, including crude samples. Commonly used quantitative methods, such as Bradford assays, can be time-consuming, require extensive amounts of manual work from the user, and are limited to working with purified samples. With Alto’s fully automated system, users can easily generate 8 standard curves and estimate concentrations of up to 40 unknown samples using the quantitation assay.
This note will demonstrate how Altoâ„¢ accurately quantifies monoclonal antibodies (mAb) in serum specific to the H3N2 hemagglutinin (HA) protein at varying concentrations. H3N2 is a variant of the H1N1 Influenza virus and uses HA, a surface glycoprotein, for viral entry into the target cell. The receptor-binding domain of HA is a popular target for antibodies due to their ability to directly inhibit binding of the virus to the host cell receptor.
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Title: Lysine-Directed Site-Selective Bioconjugation for the Creation of Radioimmunoconjugates
Over recent years, the use of monoclonal antibodies (mAbs) as tools in oncology has become increasingly common. mAbs have been harnessed to carry fluorophores, photosensitizers, and radionuclides to tumors. The remarkable selectivity and affinity of radionuclide-labelled mAbs for cancer antigens can be leveraged for imaging and radiotherapy. The publication by Sarrett et al. describes a more selective, site-specific method to build radioimmunoconjugates. In this work, the simple strategy was used to build a radio conjugate to an mAb that recognizes HER2, a protein associated with breast cancer. OpenSPR-XTâ„¢ was crucial in the validation of the newly synthesized mAbs: the use of SPR determined that the kinetics between HER2 and the newly conjugated mAbs was nearly identical to mAbs synthesized with to-date standard methods. The clever use of the Protein A coupling on OpenSPR-XTâ„¢ allowed for facile, automated testing of the antibodies to support the new technology and improve cancer technology.
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Title: Receptor-Based Peptides for Inhibition of Leukotoxin Activity
The Gram-negative bacterium Aggregatibacter actinomycetemcomitans, commonly associated with localized aggressive periodontitis (LAP), secretes an RTX (repeats-in-toxin) protein leukotoxin (LtxA) that targets human white blood cells, an interaction that is driven by its recognition of the lymphocyte function-associated antigen-1 (LFA-1) integrin. In this study, we report on the inhibition of LtxA-LFA-1 binding as an antivirulence strategy to inhibit LtxA-mediated cytotoxicity. Specifically, we designed and synthesized peptides corresponding to the reported LtxA binding domain on LFA-1 and characterized their capability to inhibit LtxA binding to LFA-1 and subsequent cytotoxic activity in human immune cells. We found that several of these peptides, corresponding to sequential β-strands in the LtxA-binding domain of LFA-1, inhibit LtxA activity, demonstrating the effectiveness of this approach. Further investigations into the mechanism by which these peptides inhibit LtxA binding to LFA-1 reveal a correlation between toxin-peptide affinity and LtxA-mediated cytotoxicity, leading to a diminished association between LtxA and LFA-1 on the cell membrane. Our results demonstrate the possibility of using target-based peptides to inhibit LtxA activity, and we expect that a similar approach could be used to hinder the activity of other RTX toxins.
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Rapid changes in influenza antiviral target proteins due to antigenic drift result in cloaking of the influenza virus from the immune system of vaccinated hosts. Hence, annual formulation updates for influenza vaccines and related antibody therapies are required to preserve immune recognition against different influenza subtypes. As such, characterizing the binding kinetics and epitope diversity of various antibodies to influenza viral antigens is essential for treating and preventing potential outbreaks. In collaboration with Sino Biological, a global leader in recombinant technology, we use Nicoya’s surface plasmon resonance (OpenSPR™) system to perform epitope characterization of several antibodies against Influenza A nucleoprotein (NP).
Read it here.
