We describe a new technique in protein synthesis that extends the existing repertoire of methods for protein modification: A chemoselective reaction that induces reactivity for a subsequent ...bioconjugation. An azide‐modified building block reacts first with an ethynylphosphonite through a Staudinger‐phosphonite reaction (SPhR) to give an ethynylphosphonamidate. The resulting electron‐deficient triple bond subsequently undergoes a cysteine‐selective reaction with proteins or antibodies. We demonstrate that ethynylphosphonamidates display excellent cysteine‐selective reactivity combined with superior stability of the thiol adducts, when compared to classical maleimide linkages. This turns our technique into a versatile and powerful tool for the facile construction of stable functional protein conjugates.
Staudinger—Ready—Go! Ethynylphosphonamidates can be chemoselectively incorporated into a given molecule through a Staudinger‐phosphonite reaction, and they react specifically with cysteine residues on proteins to give thiol adducts that are stable under physiological conditions. This enables the facile fusion of complex molecules to proteins.
TUM-VIE: The TUM Stereo Visual-Inertial Event Dataset Klenk, Simon; Chui, Jason; Demmel, Nikolaus ...
2021 IEEE/RSJ International Conference on Intelligent Robots and Systems (IROS),
2021-Sept.-27
Conference Proceeding
Event cameras are bio-inspired vision sensors which measure per pixel brightness changes. They offer numerous benefits over traditional, frame-based cameras, including low latency, high dynamic ...range, high temporal resolution and low power consumption. Thus, these sensors are suited for robotics and virtual reality applications. To foster the development of 3D perception and navigation algorithms with event cameras, we present the TUM-VIE dataset. It consists of a large variety of handheld and head-mounted sequences in indoor and outdoor environments, including rapid motion during sports and high dynamic range scenarios. The dataset contains stereo event data, stereo grayscale frames at 20Hz as well as IMU data at 200Hz. Timestamps between all sensors are synchronized in hardware. The event cameras contain a large sensor of 1280x720 pixels, which is significantly larger than the sensors used in existing stereo event datasets (at least by a factor of ten). We provide ground truth poses from a motion capture system at 120Hz during the beginning and end of each sequence, which can be used for trajectory evaluation. TUM-VIE includes challenging sequences where state-of-the art visual SLAM algorithms either fail or result in large drift. Hence, our dataset can help to push the boundary of future research on event-based visual-inertial perception algorithms.
Event cameras asynchronously capture brightness changes with low latency, high temporal resolution, and high dynamic range. However, annotation of event data is a costly and laborious process, which ...limits the use of deep learning methods for classification and other semantic tasks with the event modality. To reduce the dependency on labeled event data, we introduce Masked Event Modeling (MEM), a self-supervised framework for events. Our method pretrains a neural network on unlabeled events, which can originate from any event camera recording. Subsequently, the pretrained model is finetuned on a downstream task, leading to a consistent improvement of the task accuracy. For example, our method reaches state-of-the-art classification accuracy across three datasets, N-ImageNet, N-Cars, and N-Caltech101, increasing the top-1 accuracy of previous work by significant margins. When tested on real-world event data, MEM is even superior to supervised RGB-based pretraining. The models pretrained with MEM are also label-efficient and generalize well to the dense task of semantic image segmentation.
Multivalent interactions at biological interfaces occur frequently in nature and mediate recognition and interactions in essential physiological processes such as cell-to-cell adhesion. Multivalency ...is also a key principle that allows tight binding between pathogens and host cells during the initial stages of infection. One promising approach to prevent infection is the design of synthetic or semisynthetic multivalent binders that interfere with pathogen adhesion
. Here, we present a multivalent binder that is based on a spatially defined arrangement of ligands for the viral spike protein haemagglutinin of the influenza A virus. Complementary experimental and theoretical approaches demonstrate that bacteriophage capsids, which carry host cell haemagglutinin ligands in an arrangement matching the geometry of binding sites of the spike protein, can bind to viruses in a defined multivalent mode. These capsids cover the entire virus envelope, thus preventing its binding to the host cell as visualized by cryo-electron tomography. As a consequence, virus infection can be inhibited in vitro, ex vivo and in vivo. Such highly functionalized capsids present an alternative to strategies that target virus entry by spike-inhibiting antibodies
and peptides
or that address late steps of the viral replication cycle
.
Estimating neural radiance fields (NeRFs) from "ideal" images has been extensively studied in the computer vision community. Most approaches assume optimal illumination and slow camera motion. These ...assumptions are often violated in robotic applications, where images may contain motion blur, and the scene may not have suitable illumination. This can cause significant problems for downstream tasks such as navigation, inspection, or visualization of the scene. To alleviate these problems, we present E-NeRF, the first method which estimates a volumetric scene representation in the form of a NeRF from a fast-moving event camera. Our method can recover NeRFs during very fast motion and in high-dynamic-range conditions where frame-based approaches fail. We show that rendering high-quality frames is possible by only providing an event stream as input. Furthermore, by combining events and frames, we can estimate NeRFs of higher quality than state-of-the-art approaches under severe motion blur. We also show that combining events and frames can overcome failure cases of NeRF estimation in scenarios where only a few input views are available without requiring additional regularization.
The development of multivalent sialic acid-based inhibitors active against a variety of influenza A virus (IAV) strains has been hampered by high genetic and structural variability of the targeted ...viral hemagglutinin (HA). Here, we addressed this challenge by employing sialylated polyglycerols (PGs). Efficacy of prototypic PGs was restricted to a narrow spectrum of IAV strains. To understand this restriction, we selected IAV mutants resistant to a prototypic multivalent sialylated PG by serial passaging. Resistance mutations mapped to the receptor binding site of HA, which was accompanied by altered receptor binding profiles of mutant viruses as detected by glycan array analysis. Specifying the inhibitor functionalization to 2,6-α-sialyllactose (SL) and adjusting the linker yielded a rationally designed inhibitor covering an extended spectrum of inhibited IAV strains. These results highlight the importance of integrating virological data with chemical synthesis and structural data for the development of sialylated PGs toward broad anti-influenza compounds.
Multiple conjugation of virus-binding ligands to multivalent carriers is a prominent strategy to construct highly affine virus binders for the inhibition of viral entry into host cells. In a previous ...study, we introduced rationally designed sialic acid conjugates of bacteriophages (Qβ) that match the triangular binding site geometry on hemagglutinin spike proteins of influenza A virions, resulting in effective infection inhibition in vitro and in vivo. In this work, we demonstrate that even partially sialylated Qβ conjugates retain the inhibitory effect despite reduced activity. These observations not only support the importance of trivalent binding events in preserving high affinity, as supported by computational modeling, but also allow us to construct heterobifunctional modalities. Capsids carrying two different sialic acid ligand–linker structures showed higher viral inhibition than their monofunctional counterparts. Furthermore, capsids carrying a fluorescent dye in addition to sialic acid ligands were used to track their interaction with cells. These findings support exploring broader applications as multivalent inhibitors in the future.
A chemical route to periodic hole arrays in gold films whose holes are loaded with single gold nanoparticles is presented, paving the road to mass production of highly sensitive plasmonic sensors on ...large areas.
A chemical route to periodic hole arrays in gold films whose holes are loaded with single gold nanoparticles is presented, paving the road to mass production of highly sensitive plasmonic sensors on large areas.
In der vorliegenden Arbeit diente das Kapsid der Bakteriophage Qbeta als multivalentes Gerüst und ermöglichte die Bildung eines monodispersen multivalenten Systems, welches mit Homopropargylglycin ...als unnatürliche Aminosäure modifiziert wurde. Das so eingeführte Alkin ermöglichte kupferkatalysierte Alkin-Azid-Cycloaddition zur Anbindung von Sialinsäuregrupen. Die entsprechende Synthese der hierzu kompatiblen Azid-modifizierten Sialinsäurederivate war eine der Hauptaufgaben dieser Arbeit. Zu diesem Zweck wurde das einfach zugängliche 5-N-Acetyladamantanylthiosialosid als Glykosylierungsdonor in der alpha-selektiven Synthese von Sialosiden evaluiert. Eine effiziente Aktivierung dieses Donors wurde unter optimierten Bedingungen bei -78°C mit N-Iodsuccinimid und Trifluormethansulfonsäure erreicht, was zu hohen alpha-Selektivitäten und Gesamtausbeuten der gewünschten Sialoside führte. Insbesondere Azidoethylenglykol-verknüpfte Sialinsäuren wurden synthetisiert, die für nachfolgende Biokonjugationsreaktionen an das Qbeta-Kapsid verwendet wurden. Die so dargestellten Sialinsäure-modifizierten Qbeta-Kapsidpartikel wurden dann eingehend mit Hilfe mehrerer biophysikalischer und biologischer Tests hinsichtlich ihrer Fähigkeit an Hämagglutinin zu binden und eine Influenza-Infektion zu inhibieren charakterisiert. Niedrige nanomolare Affinitäten wurden in diesen Assays gemessen. Eine sehr effiziente Infektionshemmung in vergleichbaren Konzentrationsbereichen konnte in einem in vitro Zell-, sowie einem in vivo Maus- als auch einem menschlichen ex vivo Modellsystem beobachtet werden. Verschiedene pathologisch relevante Influenzastämme konnten über die hier vorgestellte Strategie ebenfalls gebunden werden. Die monodisperse und definierte Struktur des Qbeta-Gerüsts erlaubte es außerdem ein theoretisches Modell der zugrundeliegenden Bindungsmodi zu erstellen.
In this thesis, the bacteriophage Qbeta capsid served as a multivalent scaffold and facilitated the generation of a monodisperse multivalent system which was modified with homopropargylglycine as an unnatural amino acid. The introduced alkyne enabled copper-catalyzed alkyne-azide cycloaddition to attach sialic acid groups. The corresponding synthesis of the compatible azide-modified sialic acid derivatives was one of the main tasks of this work. For this purpose, the straightforwardly accessible 5-N-acetyladamantanyl thiosialoside was evaluated as a glycosylation donor in the alpha-selective synthesis of sialosides. Efficient activation of this donor was achieved under optimized conditions at -78°C with N-iodosuccinimide and trifluoromethanesulfonic acid which led to high alpha selectivities and overall yields of the desired sialosides. Particularly azidoethylene glycol-linked sialic acids were synthesized which were used for subsequent bioconjugation reactions to the Qbeta capsid. These synthesized sialic acid-modified Qbeta capsid particles were then thoroughly characterized by multiple biophysical and biological assays regarding their ability to bind to hemagglutinin and to inhibit influenza infection. Low nanomolar affinities were measured in these assays. A very efficient infection inhibition in a comparable concentration range was observed in in vitro cellular, in vivo mouse and ex vivo human model systems. Several pathologically relevant influenza strains could also be bound with the strategy presented here. The monodisperse and defined structure of the Qbeta scaffold additionally allowed for the establishment of a theoretical model describing the underlying binding modes.
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