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doi: 10.1038/s41591-021-01270-4. not SARS-CoV-2 except for CR3022). Structures of variable fragment (Fv) in complex with receptor binding domain name (RBD) from SARS-CoV or SARS-CoV-2 were subjected to our established in silico antibody engineering platform to improve their binding affinity to SARS-CoV-2 and developability profiles. The selected top mutations were ensembled into a focused library for each antibody for further screening. In addition, we convert the selected binders with different epitopes into the trispecific format, aiming to increase potency and to prevent mutational escape. Lastly, to avoid antibody-induced computer virus activation or enhancement, we suggest application of NNAS and DQ mutations to the Fc region to CH5424802 eliminate effector functions and lengthen half-life. Keywords: SARS-CoV-2antibody, engineeringstructure-based, engineeringtri-specific, antibodymachine learning STATEMENT OF SIGNIFICANCE Engineering SARS-CoV antibody for SARS-CoV-2 cross-reactivity is usually a potentially effective and fast way toward COVID-19 treatment. We utilized computational methods to engineer known antibodies and further formatted them into tri-specific antibody aiming for potent and broad neutralization of SARS-CoV-2. We share our proposal to contribute to the SARS-CoV-2 research community. INTRODUCTION COVID-19 cases continue to climb rapidly after causing over 160 million infections and 3.3 million deaths since the start of the outbreak. The causing computer virus, SARS-CoV-2, is usually recognized to enter human cells by binding to the angiotensin-converting enzyme 2 (ACE2) protein, following a comparable path as SARS-CoV contamination in 2003 [1C3]. However, compared to SARS, mutations in the RBD domain name in SARS-CoV-2 produce a stronger binding affinity to human ACE2 [4C7]. Due to the function of mediating cell access, the spike protein and its RBD have been the focus of drug discovery for SARS coronaviruses. To date, hundreds of new research projects are focused on exploring potential treatments, many are at the preclinical trial phase, and several have reached the administration stage. For instance, the mRNA-based vaccines developed by Moderna and Pfizer-BioNTech along with the Oxford-AstraZenecas vaccine built around the chimpanzee adenoviral vector supplemented by the SARS-CoV-2 spike protein have been authorized for emergency use. Besides vaccines, therapeutic antibodies offer additional advantages including tractable efficacy, stability and biocompatibility. Several antibody-based therapeutics to combat SAR-CoV-2 have been developed, including Regenerons REGN-CoV2 and Eli Lillys LY-CoV555. The former is usually a cocktail of two monoclonal antibodies Rabbit Polyclonal to p47 phox (mAbs), REGN10933 and REGN10987, that target different RBD regions in order to maintain its neutralizing CH5424802 activity against future mutations [8], while the latter is usually isolated from a recovering COVID-19 patient [9]. While developments of new vaccines and therapeutics have progressed rapidly, SARS-CoV-2 is usually evolving at a fast pace, if not faster, and thus poses risks and uncertainties to developed candidates and products. Several variants including K417N, E484K and N501Y mutations and deletions at positions 6970 of the RBD have been reported. One of the spike protein mutations, E484K, was suggested to hinder the neutralization effects of some polyclonal and monoclonal antibodies [10, 11]. Some early studies suggest the mRNA-based vaccines developed by Moderna and CH5424802 Pfizer-BioNTech may be less effective against the recently emerged South Africa variant [12, 13]. To increase neutralization likelihood and prevent mutational escape, application of a mixture of monoclonal antibodies, i.e. an antibody cocktail, results in stronger responses that are particularly effective against highly evolving pathogens [8]. Multispecific antibody engineering based on a combination of broadly neutralizing antibodies is usually another highly effective method to target constantly evolving viruses. This design rationale was used to generate a trispecific antibody against HIV [14]. The underlying hypothesis is usually that targeting different regions of the antigen prevents resistance and escape and further enhances cross reactivity. Similar strategy using tandem linked single domain name camelid antibodies showed significant efficacy against both influenza A and B viruses [15]. Several neutralizing mAbs targeting the spike RBD on.