Additional file 1 of Identification and application of a pair of noncompeting monoclonal antibodies broadly binding to the nucleocapsid proteins of SARS-CoV-2 variants including Omicron
Creators: name: Zhou, Bing
name: Cheng, Lin
name: Song, Shuo
name: Guo, Huimin
name: Shen, Senlin
name: Wang, Haiyan
name: Ge, Xiangyang
name: Liu, Lei
name: Ju, Bin
name: Zhang, Zheng
Publisher: name: figshare
Year: 2022
Subjects: subject: Biochemistry
subject: Pharmacology
subject: Biotechnology
subject: Chemical Sciences not elsewhere classified
subject: Immunology
subject: FOS: Clinical medicine
subject: Biological Sciences not elsewhere classified
subject: Cancer
subject: Hematology
subject: Infectious Diseases
subject: FOS: Health sciences
subject: Virology
subject: Computational Biology

description: Additional file 1. Figure S1. The isolation of a pair of noncompeting mAbs binding to SARS-CoV-2 and SARS-CoV NPs. (a) The binding activity of plasma from a convalescent COVID-19 patient to SARS-CoV-2 NP measured by ELISA. Patient: P301. HD: Healthy donor. (b) The gating strategy for isolation of SARS-CoV-2 WT NP-specific B cells by FACS. (c) The binding activities of five mAbs to NPs of SARS-CoV-2, SARS-CoV, and MERS-CoV, respectively, measured by ELISA. The pAb is used here as a positive control. The IgG1 is a negative control. (d) Competition ELISA of P301-F7 with itself and P301-H5. The IgG1 is a negative control. Figure S2. The identification of epitopes recognized by P301-F7 and P301-H5. (a) ELISA binding of P301-F7 and P301-H5 to native and denatured SARS-CoV-2 NP which was treated with the denaturing buffer containing 0.5% SDS and 40 mM DTT1 (New England Biolabs) at 100 °C for 10 mins. The pAb is used here as a positive control. VRC01 recognizing conformational epitopes of HIV-1 gp140 was served as negative control to prove that the denaturing process was efficient. (b) Construction strategies of expression vectors of the full-length and two C-terminal truncated SARS-CoV-2 NPs. NP: M1-A419. NP-T1: M1-E280. NP-T2: M1-M210. (c) The full-length and two C-terminal truncated SARS-CoV-2 NPs were expressed in 293 T cells by the transient transfection, respectively, and then detected by P301-F7 and P301-H5 using the flow cytometry analysis. The pAb is used here as a positive control. (d) The recognition epitope of P301-F7 and P301-H5 were identified by ELISA screening of peptide pools of SARS-CoV-2 NP. Figure S3. The sandwich ELISA detection of SARS-CoV and SARS-CoV-2 NPs using P301-F7 and HRP conjugated P301-H5. P301-F7 is used as a capture antibody. P301-H5-HRP is used as a detection antibody. Figure S4. The neutralizing activity of P2C-1F11 against SARS-CoV-2 WT live virus was measured by the FRNT. The IgG1 is a negative control. P301-F7-HRP is used as a detection antibody. Figure S5. Key mutations appear in the NP of SARS-CoV-2 variant viruses. Amino acid mutations of NPs in SARS-CoV-2 variants according to GISAID EpiCoV database ( https://www.gisaid.org/hcov19-mutationdashboard/ ) and outbreak.info. Figure S6. ELISA binding of P301-F7 and P301-H5 to NPs of endemic coronaviruses including HKU1, OC43, NL63 and 229E. The pAb is used here as a positive control.