Potent neutralizing antibodies against multiple epitopes on SARS-CoV-2 spike

Liu, Lihong; Wang, Pengfei; Nair, Manoj S.; Yu, Jian; Rapp, Micah; Wang, Qian; Luo, Yang; Chan, Jasper F W; Sahi, Vincent; Figueroa, Amir; Guo, Xinzheng V.; Cerutti, Gabriele; Bimela, Jude; Gorman, Jason; Zhou, Tongqing; Chen, Zhiwei; Yuen, Kwok Yung; Kwong, Peter D.; Sodroski, Joseph; Yin, Michael T.; Sheng, Zizhang; Huang, Yaoxing; Shapiro, Lawrence; Ho, David D.

doi:10.1038/s41586-020-2571-7

Cited by 1,380 publications

(1,575 citation statements)

References 45 publications

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“…The RBDs of SARS and SARS-CoV-2 are known to contain neutralising epitopes (16,(31)(32)(33)(34)(35)(36)(37)(38)(39)(40)(41)(42), and vaccines based on the RBD of SARS and SARS-CoV-2 induce strong neutralising antibodies that are protective in animal models (43)(44)(45). There is a tendency for the most potent neutralising antibodies to be those that block the binding of the RBD to its receptor ACE2 (31)(32)(33)(34)(35)(36)(37)(38)(39)(40)(41)(42)(46)(47)(48). Occasional antibodies that do not occupy the ACE2 footprint and fail to block ACE2 binding, can be almost as powerful (40), perhaps through triggering a conformational change in the spike glycoprotein that renders it non-functional (4,16).…”

Section: Neutralising Antibodiesmentioning

confidence: 99%

Breadth and function of antibody response to acute SARS-CoV-2 infection in humans

Huang

Tan

Chen

et al. 2020

Preprint

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Plasmablast responses and derived IgG monoclonal antibodies (MAbs) have been analysed in three COVID-19 patients. An average of 13.7% and 13.0% of plasmablast-derived IgG MAbs were reactive with virus spike glycoprotein or nucleocapsid, respectively. Of thirty-two antibodies specific for the spike glycoprotein, ten recognised the receptor-binding domain (RBD), thirteen were specific for non-RBD epitopes on the S1 subunit, and nine recognised the S2 subunit. A subset of anti-spike antibodies (10 of 32) cross-reacted with other betacoronaviruses tested, five targeted the non-RBD S1, and five targeted the S2 subunit. Of the plasmablast-derived MAbs reacting with nucleocapsid, over half of them (19 of 35) cross-reacted with other betacoronaviruses tested. The cross-reactive plasmablast-derived antibodies harboured extensive somatic mutations, indicative of an expansion of memory B cells upon SARS-CoV-2 infection. We identified 14 of 32 anti-spike MAbs that neutralised SARS-CoV-2 in independent assays at ≤ 133 nM (20 μg/ml) (five of 10 anti-RBD, three of 13 anti-non-RBD S1 subunit, six of nine anti-S2 subunit). Six of 10 anti-RBD MAbs showed evidence of blockade of ACE2 binding to RBD, and five of six of these were neutralising. Non-competing pairs of neutralising antibodies were identified, which offer potential templates for the development of prophylactic and therapeutic agents against SARS-CoV-2.

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Section: Neutralising Antibodiesmentioning

confidence: 99%

Breadth and function of antibody response to acute SARS-CoV-2 infection in humans

Huang

Tan

Chen

et al. 2020

Preprint

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“…Overall, these structural analyses indicate that RBD rotational flexibility and acquisition of quaternary interactions can play an important role in CR3022 interaction with the S protein. CR3022 adds to the growing list of neutralization antibodies that can utilize quaternary interactions for binding to the S protein [12,36].…”

Section: Rbd Flexibility and Quaternary Interactions In Cr3022-boundmentioning

confidence: 99%

“…The major antigen of coronavirus is the spike (S) glycoprotein, which is expressed as a homotrimer on the virus surface. Since the S protein is essential for virus entry through engaging the host receptor and mediating virus-host membrane fusion, many antibodies to the S protein are neutralizing [1][2][3][4][5][6][7][8][9][10][11][12]. The S proteins of SARS-CoV-2 and SARS-CoV, which caused a global outbreak in 2003, have an amino-acid sequence identity of around 77% [13] that leads to differences in antigenicity in serology studies [14,15].…”

Section: Introductionmentioning

confidence: 99%

“…The CR3022 epitope is exposed only when the RBD is in the "up" but not the "down" conformation on the S protein [20]. A few SARS-CoV-2 antibodies from patients have also recently been shown to target the CR3022 epitope [12,17,21], suggesting that it is an important site of vulnerability for the antibody response in SARS-CoV-2 infection. Out of 28 residues in the CR3022 epitope, 24 are conserved between SARS-CoV-2 and SARS-CoV, explaining the cross-reactive binding of CR3022.…”

Section: Introductionmentioning

confidence: 99%

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A natural mutation between SARS-CoV-2 and SARS-CoV determines neutralization by a cross-reactive antibody

Yuan

Bangaru

et al. 2020

Preprint

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Epitopes that are conserved among SARS-like coronaviruses are attractive targets for design of cross-reactive vaccines and therapeutics. CR3022 is a SARS-CoV neutralizing antibody to a highly conserved epitope on the receptor binding domain (RBD) on the spike protein that can cross-react with SARS-CoV-2, but with lower affinity. Using x-ray crystallography, mutagenesis, and binding experiments, we illustrate that of four amino acid differences in the CR3022 epitope between SARS-CoV-2 and SARS-CoV, a single mutation P384A fully determines the affinity difference. CR3022 does not neutralize SARS-CoV-2, but the increased affinity to SARS-CoV-2 P384A mutant now enables neutralization with a similar potency to SARS-CoV. We further investigated CR3022 interaction with the SARS-CoV spike protein by negative-stain EM and cryo-EM. Three CR3022 Fabs bind per trimer with the RBD observed in different up-conformations due to considerable flexibility of the RBD. In one of these conformations, quaternary interactions are made by CR3022 to the N-terminal domain (NTD) of an adjacent subunit. Overall, this study provides insights into antigenic variation and potential for cross-neutralizing epitopes on SARS-like viruses.

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“…In Spike-2P immunized animals titers were more variable than in mRBD immunized animals though the difference in neutralization titers did not approach statistical significance. A potential advantage of using the spike as an immunogen is that it contains neutralization epitopes outside the RBD, including in the NTD 32 . We therefore probed the Spike-2P sera for NTD titers using a mammalian cell expressed NTD construct.…”

Section: Addavax™ Adjuvanted Rbd Elicits Neutralizing Antibodies In Gmentioning

confidence: 99%

Design of a highly thermotolerant, immunogenic SARS-CoV-2 spike fragment immunogen

Malladi

Singh

Pandey

et al. 2020

Preprint

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Virtually all SARS-CoV-2 vaccines currently in clinical testing are stored in a refrigerated or frozen state prior to use. This is a major impediment to deployment in resource-poor settings. Several use viral vectors or mRNA. In contrast to protein subunit vaccines, there is limited manufacturing expertise for these novel, nucleic acid based modalities, especially in the developing world. Neutralizing antibodies, the clearest known correlate of protection against SARS-CoV-2, are primarily directed against the Receptor Binding Domain (RBD) of the viral spike protein. We describe a monomeric, glycan engineered RBD protein fragment that is expressed at a purified yield of 200mg/L in unoptimized, mammalian cell culture and in contrast to a stabilized spike ectodomain, is tolerant of exposure to temperatures as high as 100°C when lyophilized, and upto 70°C in solution. In prime:boost guinea pig immunizations, when formulated with the MF59 like adjuvant AddaVax™, the RBD derivative elicited neutralizing antibodies with an endpoint geometric mean titer of ~415 against replicative virus, comparing favourably with several vaccine formulations currently in the clinic. These features of high yield, extreme thermotolerance and satisfactory immunogenicity suggest that such RBD subunit vaccine formulations hold great promise to combat COVID-19.

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Potent neutralizing antibodies against multiple epitopes on SARS-CoV-2 spike

Cited by 1,380 publications

References 45 publications

Breadth and function of antibody response to acute SARS-CoV-2 infection in humans

Breadth and function of antibody response to acute SARS-CoV-2 infection in humans

A natural mutation between SARS-CoV-2 and SARS-CoV determines neutralization by a cross-reactive antibody

Design of a highly thermotolerant, immunogenic SARS-CoV-2 spike fragment immunogen

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