key: cord-0694296-fyuu69f8 authors: Makdasi, Efi; Zvi, Anat; Alcalay, Ron; Noy-Porat, Tal; Peretz, Eldar; Mechaly, Adva; Levy, Yinon; Epstein, Eyal; Chitlaru, Theodor; Tennenhouse, Ariel; Aftalion, Moshe; Gur, David; Paran, Nir; Tamir, Hadas; Zimhony, Oren; Weiss, Shay; Mandelboim, Michal; Mendelson, Ella; Zuckerman, Neta; Nemet, Ital; Kliker, Limor; Yitzhaki, Shmuel; Shapira, Shmuel C.; Israely, Tomer; Fleishman, Sarel J.; Mazor, Ohad; Rosenfeld, Ronit title: The neutralization potency of anti-SARS-CoV-2 therapeutic human monoclonal antibodies is retained against viral variants date: 2021-08-21 journal: Cell Rep DOI: 10.1016/j.celrep.2021.109679 sha: 912bddcc75d79a306b9eafaff756cd014601d7b2 doc_id: 694296 cord_uid: fyuu69f8 A wide range of SARS-CoV-2 neutralizing monoclonal antibodies (mAbs) were reported, most of which target the spike glycoprotein. Therapeutic implementation of these antibodies has been challenged by emerging SARS-CoV-2 variants harboring mutated spike versions. Consequently, re-assessment of previously identified mAbs is of high priority. Four previously selected mAbs targeting non-overlapping epitopes are now evaluated for binding potency to mutated RBD versions, reported to mediate escape from antibody neutralization. In vitro neutralization potencies of these mAbs, and two NTD-specific mAbs, are evaluated against two frequent SARS-CoV-2 variants of concern, the B.1.1.7 Alpha, and the B.1.351 Beta. Furthermore, we demonstrate therapeutic potential of three selected mAbs by treatment of K18-hACE2 transgenic mice two days post infection with each virus variant. Thus, despite the accumulation of spike mutations, the highly potent MD65 and BL6 mAbs retain their ability to bind the prevalent viral mutants, effectively protecting against B.1.1.7 and B.1.351 variants. A wide range of SARS-CoV-2 neutralizing monoclonal antibodies (mAbs) were 25 reported, most of which target the spike glycoprotein. Therapeutic implementation of 26 these antibodies has been challenged by emerging SARS-CoV-2 variants harboring 27 mutated spike versions. Consequently, re-assessment of previously identified mAbs is of 28 high priority. Four previously selected mAbs targeting non-overlapping epitopes are now 29 evaluated for binding potency to mutated RBD versions, reported to mediate escape from 30 antibody neutralization. In vitro neutralization potencies of these mAbs, and two NTD-31 specific mAbs, are evaluated against two frequent SARS-CoV-2 variants of concern, the 32 B.1.1.7 Alpha, and the B.1.351 Beta. Furthermore, we demonstrate therapeutic potential Prior to its global expansion, SARS-CoV-2 was expected to exhibit relatively low 58 mutation rates, as compared to other RNA viruses since its genome encodes a 59 proofreading exoribonuclease (Robson et al., 2020) . Nevertheless, the rapid global spread now represented by SARS-CoV-2 emerged genetic variants which spread worldwide. 128 We previously reported the isolation of RBD-and NTD-specific highly-neutralizing and Y453F (schematically described in Figure S3A ; for details, see https://outbreak.info). Biolayer interferometry (BLI) analysis was applied to evaluate the ability of the RBD- Thus, in line with the individual mutation binding results (see Figure 1E for LY-CoV555 antibody with respect to the three tested viral strains are tabulated in Figure 4H . Table S1 . The following His-tagged recombinant proteins were purchased from Sino Biologicals: Table S1 . Area under curve (AUC) was calculated for each binding curve presented in Figure 3 . Results are presented in Table S1 . For all experiments, exact value and meaning of n are presented in the figure legends. For antibody structure modeling, the amino acid sequence of the variable domain of 608 MD65 was submitted to the AbPredict2 webserver which is available freely for non-609 commercial use (http://AbPredict.weizmann.ac.il). 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