Rational development of a human antibody cocktail that deploys multiple functions to confer Pan-SARS-CoVs protection

AbstractStructural principles underlying the composition and synergistic mechanisms of protective monoclonal antibody cocktails are poorly defined. Here, we exploited antibody cooperativity to develop a therapeutic antibody cocktail against SARS-CoV-2. On the basis of our previously identified humanized cross-neutralizing antibody H014, we systematically analyzed a fully human naive antibody library and rationally identified a potent neutralizing antibody partner, P17, which confers effective protection in animal model. Cryo-EM studies dissected the nature of the P17 epitope, which is SARS-CoV-2 specific and distinctly different from that of H014. High-resolution structure of the SARS-CoV-2 spike in complex with H014 and P17, together with functional investigations revealed that in a two-antibody cocktail, synergistic neutralization was achieved by S1 shielding and conformational locking, thereby blocking receptor attachment and viral membrane fusion, conferring high potency as well as robustness against viral mutation escape. Furthermore, cluster analysis identified a hypothetical 3rd antibody partner for further reinforcing the cocktail as pan-SARS-CoVs therapeutics.

Download Full-text

Structure of a germline-like human antibody defines a neutralizing epitope on the SARS-CoV-2 spike NTD

10.1101/2021.07.08.451649 ◽

2021 ◽

Author(s):

Clara Gilda Altomare ◽

Daniel Cole Adelsberg ◽

Juan Manuel Carreno ◽

Iden Avery Sapse ◽

Fatima Amanat ◽

...

Keyword(s):

Arms Race ◽

Human Antibody ◽

Neutralizing Epitope ◽

Vaccine Immunogen ◽

Direct Competition ◽

High Resolution Structure ◽

Immunogen Design ◽

Insight Into ◽

Resolution Structure

Structural characterization of infection- and vaccination-elicited antibodies in complex with antigen provides insight into the evolutionary arms race between the host and the pathogen and informs rational vaccine immunogen design. We isolated a germline-like monoclonal antibody (mAb) from plasmablasts activated upon mRNA vaccination against SARS-CoV-2 and determined its structure in complex with the spike glycoprotein by cryo-EM. We show that the mAb engages a previously uncharacterized neutralizing epitope on the spike N-terminal domain (NTD). The high-resolution structure reveals details of the intermolecular interactions and shows that the mAb inserts its HCDR3 loop into a hydrophobic NTD cavity previously shown to bind a heme metabolite, biliverdin. We demonstrate direct competition with biliverdin and that - because of the conserved nature of the epitope - the mAb maintains binding to viral variants B.1.1.7 and B.1.351. Our study illustrates the feasibility of targeting the NTD to achieve broad neutralization against SARS-CoV-2 variants.

Download Full-text

High-resolution structure determination by ALCHEMI

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100074707 ◽

1983 ◽

Vol 41 ◽

pp. 178-181 ◽

Cited By ~ 1

Author(s):

Peter G. Self ◽

Peter R. Buseck

Keyword(s):

Site Occupancy ◽

Real Space ◽

Unit Cell ◽

Bragg Angle ◽

Electron Current ◽

High Resolution Structure ◽

Beam Incident ◽

Crystallographic Planes ◽

Electron Beam Incident ◽

Resolution Structure

ALCHEMI (Atom Location by CHanneling Enhanced Microanalysis) enables the site occupancy of atoms in single crystals to be determined. In this article the fundamentals of the method for both EDS and EELS will be discussed. Unlike HRTEM, ALCHEMI does not place stringent resolution requirements on the microscope and, because EDS clearly distinguishes between elements of similar atomic number, it can offer some advantages over HRTEM. It does however, place certain constraints on the crystal. These constraints are: a) the sites of interest must lie on alternate crystallographic planes, b) the projected charge density on the alternate planes must be significantly different, and c) there must be at least one atomic species that lies solely on one of the planes.An electron beam incident on a crystal undergoes elastic scattering; in reciprocal space this is seen as a diffraction pattern and in real space this is a modulation of the electron current across the unit cell. When diffraction is strong (i.e., when the crystal is oriented near to the Bragg angle of a low-order reflection) the electron current at one point in the unit cell will differ significantly from that at another point.

Download Full-text

Cathodoluminescence of Catalyst Crystallites

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100055667 ◽

1982 ◽

Vol 40 ◽

pp. 664-665

Author(s):

E.D. Boyes ◽

P.L. Gai ◽

D.B. Darby ◽

C. Warwick

Keyword(s):

Defect Density ◽

Chemical Properties ◽

Operating Conditions ◽

Semiconductor Materials ◽

Environmental Cell ◽

High Resolution Structure ◽

Commercially Important ◽

Crystallographic Defects ◽

Resolution Structure

The extended crystallographic defects introduced into some oxide catalysts under operating conditions may be a consequence and accommodation of the changes produced by the catalytic activity, rather than always being the origin of the reactivity. Operation without such defects has been established for the commercially important tellurium molybdate system. in addition it is clear that the point defect density and the electronic structure can both have a significant influence on the chemical properties and hence on the effectiveness (activity and selectivity) of the material as a catalyst. SEM/probe techniques more commonly applied to semiconductor materials, have been investigated to supplement the information obtained from in-situ environmental cell HVEM, ultra-high resolution structure imaging and more conventional AEM and EPMA chemical microanalysis.

Download Full-text