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CUSABIO induced an immune response by immunizing a mouse with a human SARS-CoV-2 spike glycoprotein (S) (16-685aa). B cells were then isolated from the immunized mouse and fused with myeloma cells, resulting in the formation of hybridoma cells. From the screened hybridoma cells, a single clone that produces the desired human SARS-CoV-2 S-specific antibody was selected. RNA was extracted from the selected hybridoma cells and the variable regions of the human SARS-CoV-2 S antibody were isolated and amplified using reverse transcription PCR. Insert the DNA sequence encoding the mouse single-chain variable fragment (scFv) into an expression vector and introduce the DNA sequence encoding the human IgG1 Fc region into the same expression vector, downstream of the mouse scFv sequence, creating a fusion construct that consists of the scFv followed by the Fc region. The recombinant vector was transfected into a host cell line for expression. The S recombinant monoclonal antibodies were purified from the cell culture supernatant using affinity chromatography. The binding specificity and affinity of the S recombinant monoclonal antibody were confirmed using various applications including ELISA, GICA, and neutralizing. This antibody specifically recognizes the human SARS-CoV-2 S protein.
Applications : Fabrication of MO CM Biochips
Sample type: magneto-optical biochip
Review: the antibody of spike glycoprotein (anti-S, 10 μg/mL, 40 μL) was added and conjugated with GA. Te absorption peaks of the biochip after the fabrication process of Au nanostructure, APTMS, GA and BSA are 532 nm, 537 nm, 538 nm and 547 nm, respectively.