Journal: Nat Commun / Year: 2022 Title: Engineering SARS-CoV-2 specific cocktail antibodies into a bispecific format improves neutralizing potency and breadth. Authors: Zhiqiang Ku / Xuping Xie / Jianqing Lin / Peng Gao / Bin Wu / Abbas El Sahili / Hang Su / Yang Liu / Xiaohua Ye / Eddie Yongjun Tan / Xin Li / Xuejun Fan / Boon Chong Goh / Wei Xiong / ...Authors: Zhiqiang Ku / Xuping Xie / Jianqing Lin / Peng Gao / Bin Wu / Abbas El Sahili / Hang Su / Yang Liu / Xiaohua Ye / Eddie Yongjun Tan / Xin Li / Xuejun Fan / Boon Chong Goh / Wei Xiong / Hannah Boyd / Antonio E Muruato / Hui Deng / Hongjie Xia / Jing Zou / Birte K Kalveram / Vineet D Menachery / Ningyan Zhang / Julien Lescar / Pei-Yong Shi / Zhiqiang An / Abstract: One major limitation of neutralizing antibody-based COVID-19 therapy is the requirement of costly cocktails to reduce emergence of antibody resistance. Here we engineer two bispecific antibodies ...One major limitation of neutralizing antibody-based COVID-19 therapy is the requirement of costly cocktails to reduce emergence of antibody resistance. Here we engineer two bispecific antibodies (bsAbs) using distinct designs and compared them with parental antibodies and their cocktail. Single molecules of both bsAbs block the two epitopes targeted by parental antibodies on the receptor-binding domain (RBD). However, bsAb with the IgG-(scFv) design (14-H-06) but not the CrossMAb design (14-crs-06) shows increased antigen-binding and virus-neutralizing activities against multiple SARS-CoV-2 variants as well as increased breadth of neutralizing activity compared to the cocktail. X-ray crystallography and cryo-EM reveal distinct binding models for individual cocktail antibodies, and computational simulations suggest higher inter-spike crosslinking potentials by 14-H-06 than 14-crs-06. In mouse models of infections by SARS-CoV-2 and multiple variants, 14-H-06 exhibits higher or equivalent therapeutic efficacy than the cocktail. Rationally engineered bsAbs represent a cost-effective alternative to antibody cocktails and a promising strategy to improve potency and breadth.
Complex: receptor binding domain of wild-type spike
Protein or peptide: Spike protein S1
Complex: Fab14
Protein or peptide: Fab14 light chain
Protein or peptide: Fab14 heavy chain
Ligand: 2-acetamido-2-deoxy-beta-D-glucopyranose
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Supramolecule #1: RBD bound to Fab14
Supramolecule
Name: RBD bound to Fab14 / type: complex / Chimera: Yes / ID: 1 / Parent: 0 / Macromolecule list: #1-#3 Details: Fab14 was generated by cleaving IgG14 with papain. RBD was obtained by first expressing a MBP-RBD fusion construct, followed by removal of MBP tag using TEV cleavage. A TEV cleavage site was ...Details: Fab14 was generated by cleaving IgG14 with papain. RBD was obtained by first expressing a MBP-RBD fusion construct, followed by removal of MBP tag using TEV cleavage. A TEV cleavage site was present between MBP and RBD. RBD-Fab14 complex was prepared by mixing both in equal molar ratio, followed by purification via SEC.
Recombinant expression
Organism: Homo sapiens (human)
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Supramolecule #2: receptor binding domain of wild-type spike
Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Number grids imaged: 1 / Number real images: 3410 / Average exposure time: 5.3 sec. / Average electron dose: 45.56 e/Å2 Details: Images were collected in movie-mode: 40 frames per 5.3 seconds.
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
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Image processing
Particle selection
Number selected: 5334997 Details: auto-picked and extracted in REION, with a box size of 256x256 pixels
CTF correction
Software - Name: RELION (ver. 3.0)
Initial angle assignment
Type: NOT APPLICABLE
Final angle assignment
Type: NOT APPLICABLE Details: Cryosparc autogenerated model based on 2D selected particles
Final reconstruction
Number classes used: 1 / Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 7.3 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC / Number images used: 117831
In the structure databanks used in Yorodumi, some data are registered as the other names, "COVID-19 virus" and "2019-nCoV". Here are the details of the virus and the list of structure data.
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