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Preparation and characterization of monoclonal antibodies <t>against</t> <t>GoAstV-2</t> VP27 protein. (A) Schematic of the immunization of mice with purified recombinant VP27 protein. The red blood drop symbols represent the time points when serum samples were collected, the notation for splenocytes indicates the time point at which the spleens were aseptically harvested. (B) The serum titers of four mice were measured by indirect ELISA. Control: negative mice immunized with <t>PBS.</t> (C) The reactivity of mAb 11-39B was tested in VP27 protein-based indirect ELISA. The cell supernatant of hybridoma clones 11-39B was used as the primary antibodies, the SP2/0 cell supernatant was used as the negative control. (D) Identification of the subtypes of mAb 11-39B. (E) Antibodies were purified using a protein G column and analyzed by SDS-PAGE. M, protein marker; Lane 1, crude ascites; Lane 2, loaded fraction; Lanes 3-4, wash fraction; Lanes 5-6, eluent fraction. (F) Measurement of the titers of ascites mAb 11-39B by VP27 protein-based indirect ELISA.
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Preparation and characterization of monoclonal antibodies against GoAstV-2 VP27 protein. (A) Schematic of the immunization of mice with purified recombinant VP27 protein. The red blood drop symbols represent the time points when serum samples were collected, the notation for splenocytes indicates the time point at which the spleens were aseptically harvested. (B) The serum titers of four mice were measured by indirect ELISA. Control: negative mice immunized with PBS. (C) The reactivity of mAb 11-39B was tested in VP27 protein-based indirect ELISA. The cell supernatant of hybridoma clones 11-39B was used as the primary antibodies, the SP2/0 cell supernatant was used as the negative control. (D) Identification of the subtypes of mAb 11-39B. (E) Antibodies were purified using a protein G column and analyzed by SDS-PAGE. M, protein marker; Lane 1, crude ascites; Lane 2, loaded fraction; Lanes 3-4, wash fraction; Lanes 5-6, eluent fraction. (F) Measurement of the titers of ascites mAb 11-39B by VP27 protein-based indirect ELISA.

Journal: Poultry Science

Article Title: Development of monoclonal antibodies for GoAstV-2 VP27 protein and precise mapping of linear antigenic epitopes

doi: 10.1016/j.psj.2026.106535

Figure Lengend Snippet: Preparation and characterization of monoclonal antibodies against GoAstV-2 VP27 protein. (A) Schematic of the immunization of mice with purified recombinant VP27 protein. The red blood drop symbols represent the time points when serum samples were collected, the notation for splenocytes indicates the time point at which the spleens were aseptically harvested. (B) The serum titers of four mice were measured by indirect ELISA. Control: negative mice immunized with PBS. (C) The reactivity of mAb 11-39B was tested in VP27 protein-based indirect ELISA. The cell supernatant of hybridoma clones 11-39B was used as the primary antibodies, the SP2/0 cell supernatant was used as the negative control. (D) Identification of the subtypes of mAb 11-39B. (E) Antibodies were purified using a protein G column and analyzed by SDS-PAGE. M, protein marker; Lane 1, crude ascites; Lane 2, loaded fraction; Lanes 3-4, wash fraction; Lanes 5-6, eluent fraction. (F) Measurement of the titers of ascites mAb 11-39B by VP27 protein-based indirect ELISA.

Article Snippet: Simultaneously, the mock-infected group was inoculated with 1 mL sterile PBS; the virus-infected group was inoculated with 0.5 mL of GoAstV-2 solution (10 5.167 ELD 50 /0.5 mL) and 0.5 mL sterile PBS containing 200 μg/mL mouse IgG control (Thermo Fisher Scientific, USA).

Techniques: Bioprocessing, Purification, Recombinant, Indirect ELISA, Control, Clone Assay, Negative Control, SDS Page, Marker

Journal: STAR Protocols

Article Title: Protocol to isolate broadly neutralizing monoclonal antibodies against SARS-CoV-2 from human B cells

doi: 10.1016/j.xpro.2025.104275

Figure Lengend Snippet:

Article Snippet: Note: The purpose of this step is to change the buffer to 1×PBS. k. Determine the protein concentration using a nanodrop spectrophotometer. l. Analyze the purified protein and the samples collected in Step a and Step f by SDS-PAGE.

Techniques: Virus, Recombinant, Sterility, Saline, Magnetic Beads, Expressing, Reverse Transcription, Cloning, Luciferase, Plasmid Preparation, Purification, DNA Purification, Software, Cell Culture, Binding Assay, Enzyme-linked Immunosorbent Assay, Adhesive