Cyanovirin-N Binds Viral Envelope Proteins at the Low-Affinity Carbohydrate Binding Site without Direct Virus Neutralization Ability

Glycan-targeting antibodies and pseudo-antibodies have been extensively studied for their stoichiometry, avidity, and their interactions with the rapidly modifying glycan shield of influenza A. Broadly neutralizing antiviral agents bind in the same order when they neutralize enveloped viruses regardless of the location of epitopes to the host receptor binding site. Herein, we investigated the binding of cyanovirin-N (CV–N) to surface-expressed glycoproteins such as those of human immunodeficiency virus (HIV) gp120, hemagglutinin (HA), and Ebola (GP)1,2 and compared their binding affinities with the binding response to the trimer-folded gp140 using surface plasmon resonance (SPR). Binding-site knockout variants of an engineered dimeric CV–N molecule (CVN2) revealed a binding affinity that correlated with the number of (high-) affinity binding sites. Binding curves were specific for the interaction with N-linked glycans upon binding with two low-affinity carbohydrate binding sites. This biologically active assembly of a domain-swapped CVN2, or monomeric CV–N, bound to HA with a maximum KD of 2.7 nM. All three envelope spike proteins were recognized at a nanomolar KD, whereas binding to HIV neutralizing 2G12 by targeting HA and Ebola GP1,2 was measured in the µM range and specific for the bivalent binding scheme in SPR. In conclusion, invariant structural protein patterns provide a substrate for affinity maturation in the membrane-anchored HA regions, as well as the glycan shield on the membrane-distal HA top part. They can also induce high-affinity binding in antiviral CV–N to HA at two sites, and CVN2 binding is achieved at low-affinity binding sites

Ultra-High Field NMR and MRI—The Role of Magnet Technology to Increase Sensitivity and Specificity

Starting with post-war developments in nuclear magnetic resonance (NMR) a race for stronger and stronger magnetic fields has begun in the 1950s to overcome the inherently low sensitivity of this promising method. Further challenges were larger magnet bores to accommodate small animals and eventually humans. Initially, resistive electromagnets with small pole distances, or sample volumes, and field strengths up to 2.35 T (or 100 MHz H frequency) were used in applications in physics, chemistry, and material science. This was followed by stronger and more stable (Nb-Ti based) superconducting magnet technology typically implemented first for small-bore systems in analytical chemistry, biochemistry and structural biology, and eventually allowing larger horizontal-bore magnets with diameters large enough to fit small laboratory animals. By the end of the 1970s, first low-field resistive magnets big enough to accommodate humans were developed and superconducting whole-body systems followed. Currently, cutting-edge analytical NMR systems are available at proton frequencies up to 1 GHz (23.5 T) based on NbSn at 1.9 K. A new 1.2 GHz system (28 T) at 1.9 K, operating in persistent mode but using a combination of low and high temperature multi-filament superconductors is to be released. Preclinical instruments range from small-bore animal systems with typically 600800 MHz (14.118.8 T) up to 900 MHz (21 T) at 1.9 K. Human whole-body MRI systems currently operate up to 10.5 T. Hybrid combined superconducting and resistive electromagnets with even higher field strength of 45 T dc and 100 T pulsed, are available for material research, of course with smaller free bore diameters. This rather costly development toward higher and higher field strength is a consequence of the inherently low and, thus, urgently needed sensitivity in all NMR experiments. This review particularly describes and compares the developments in superconducting magnet technology and, thus, sensitivity in three fields of research: analytical NMR, biomedical and preclinical research, and human MRI and MRS, highlighting important steps and innovations. In addition, we summarize our knowledge on safety issues. An outlook into even stronger magnetic fields using different superconducting materials and/or hybrid magnet designs is presented.(VLID)486222

Nuclear Magnetic Resonance Spectroscopy

Department of Structural and Computational Biology, MFP

Nuclear Magnetic Resonance Spectroscopy

Department für Strukturbiologie und Computational Biolog

Ultra-High Field NMR and MRI—The Role of Magnet Technology to Increase Sensitivity and Specificity

“History, of course, is difficult to write, if for no other reason, than that it has so many players and so many authors.” – P. J. Keating (former Australian Prime Minister)Starting with post-war developments in nuclear magnetic resonance (NMR) a race for stronger and stronger magnetic fields has begun in the 1950s to overcome the inherently low sensitivity of this promising method. Further challenges were larger magnet bores to accommodate small animals and eventually humans. Initially, resistive electromagnets with small pole distances, or sample volumes, and field strengths up to 2.35 T (or 100 MHz 1H frequency) were used in applications in physics, chemistry, and material science. This was followed by stronger and more stable (Nb-Ti based) superconducting magnet technology typically implemented first for small-bore systems in analytical chemistry, biochemistry and structural biology, and eventually allowing larger horizontal-bore magnets with diameters large enough to fit small laboratory animals. By the end of the 1970s, first low-field resistive magnets big enough to accommodate humans were developed and superconducting whole-body systems followed. Currently, cutting-edge analytical NMR systems are available at proton frequencies up to 1 GHz (23.5 T) based on Nb3Sn at 1.9 K. A new 1.2 GHz system (28 T) at 1.9 K, operating in persistent mode but using a combination of low and high temperature multi-filament superconductors is to be released. Preclinical instruments range from small-bore animal systems with typically 600–800 MHz (14.1–18.8 T) up to 900 MHz (21 T) at 1.9 K. Human whole-body MRI systems currently operate up to 10.5 T. Hybrid combined superconducting and resistive electromagnets with even higher field strength of 45 T dc and 100 T pulsed, are available for material research, of course with smaller free bore diameters. This rather costly development toward higher and higher field strength is a consequence of the inherently low and, thus, urgently needed sensitivity in all NMR experiments. This review particularly describes and compares the developments in superconducting magnet technology and, thus, sensitivity in three fields of research: analytical NMR, biomedical and preclinical research, and human MRI and MRS, highlighting important steps and innovations. In addition, we summarize our knowledge on safety issues. An outlook into even stronger magnetic fields using different superconducting materials and/or hybrid magnet designs is presented