A broadly neutralizing humanized ACE2-targeting antibody against SARS-CoV-2 variants

Abstract

The successive emergences and accelerating spread of novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) lineages and evolved resistance to some ongoing clinical therapeutics increase the risks associated with the coronavirus disease 2019 (COVID-19) pandemic. An urgent intervention for broadly effective therapies to limit the morbidity and mortality of COVID-19 and future transmission events from SARS-related coronaviruses (SARSr-CoVs) is needed. Here, we isolate and humanize an angiotensin-converting enzyme-2 (ACE2)-blocking monoclonal antibody (MAb), named h11B11, which exhibits potent inhibitory activity against SARS-CoV and circulating global SARS-CoV-2 lineages. When administered therapeutically or prophylactically in the hACE2 mouse model, h11B11 alleviates and prevents SARS-CoV-2 replication and virus-induced pathological syndromes. No significant changes in blood pressure and hematology chemistry toxicology were observed after injections of multiple high dosages of h11B11 in cynomolgus monkeys. Analysis of the structures of the h11B11/ACE2 and receptor-binding domain (RBD)/ACE2 complexes shows hindrance and epitope competition of the MAb and RBD for the receptor. Together, these results suggest h11B11 as a potential therapeutic countermeasure against SARS-CoV, SARS-CoV-2, and escape variants.

Topical TMPRSS2 inhibition prevents SARS-CoV-2 infection in differentiated human airway cultures

“…Each Transwell was washed three times with PBS and fixed using 4% PFA for 15 min at room temperature before antibody staining. The following antibodies were used: anti-ACE2 antibody (Anti-ACE2 antibody)—21115-1-AP; ProteinTech ( Du et al, 2021 ); SARS-CoV/SARS-CoV-2 (COVID-19) spike antibody [1A9] (GTX632604; GeneTex); anti-SARS-CoV-2 nucleoprotein antibody (DA114; MRC-PPU); acetylated tubulin (T7451; Sigma-Aldrich); Muc5AC (MA5-12178; Invitrogen); Hoechst dye solution (100 μg/ml) was used for nuclei staining. Confocal images were taken using Nikon Confocal Microscopes C2, magnification 40× oil.…”

Advances in the development of therapeutic strategies against COVID-19 and perspectives in the drug design for emerging SARS-CoV-2 variants

“…In a study, using protein engineering technology, the self-assembled tetramerization domain from p53 protein produces a super tetravalent form of ACE2 coupled to the Fc region of human immunoglobulin γ1, the high-molecular-weight Quad protein (ACE2-Fc-TD) retains the binding to the ACE2 and its binding SARS-CoV-2 S protein and can form a complex with the S protein and antiviral antibodies as a bait protein [54] . Similarly, antibodies that bind to viral RBD epitopes (such as REGE-CoV) [55] , [56] or that target the ACE2 receptor (such as h11B11) [57] can also effectively prevent and reduce the symptoms of viral infections. The viral load in the SARS-CoV-2 infected animals is significantly reduced after the single-dose h11B11 treatment, while the animals showed less interstitial pneumonia and limited pathological features under the preventive treatment conditions [57] , which makes it a potential therapeutical agent in COVID-19.…”

Close relatives of MERS-CoV in bats use ACE2 as their functional receptors

“…For Dual split protein (DSP) based cell-cell fusion assay, the dual reporter split proteins were expressed by pLVX-IRES-puro vector expressing the RLucaa1-155-GFP1-7(aa1-157) and GFP8-11(aa158-231)-RLuc-aa156-311 plasmids, which were constructed in the lab based on a previously study 32, 45 . The plasmids expressing the codon-optimized anti-ACE2 antibody (H11B11; GenBank accession codes MZ514137 and MZ514138) were constructed by inserting the heavy-chain and light-chain coding sequences into the pCAGGS vector with N-terminal CD5 leader sequences, respectively 34 . For anti-MERS-CoV nanobody-hFc fusion proteins, nanobody coding sequences were synthesized and cloned into the pCAGGS vector with N-terminal CD5 leader sequences and C-terminal hFc tags 35 .…”

“…In addition, the NeoCoV-T510F can enter the human colon cell line Caco-2 with much higher efficiency than wild-type NeoCoV. It enters the Caco-2 exclusively through ACE2 as the infection can be neutralized by an ACE2-targeting neutralizing antibody H11B11 34 (Fig. 4l).…”