Fig. 3.

Approaches for the development of therapeutic antibodies. a The traditional mouse hybridoma technique starts by immunization of mice with desired antigens to trigger an immune response. Harvested splenocytes are fused with myeloma cells to produce hybridoma cells that persistently secrete antibodies. After the screening, selected leads are used to generate chimeric or humanized antibodies. b Phage display. A human phage-displayed human antibody library is used to select antigens of interest. After 3–5 rounds of biopanning, immuno-positive phage clones are screened by ELISA; then DNA sequences are analyzed to construct and express human IgGs. c Transgenic mouse. Similar to the mouse hybridoma technique or single B cell methods. d The single B cell technique. From infected or vaccinated donors, PBMCs are prepared for isolation of suitable B cells by flow cytometry. Following the RT-PCR, V_H and V_L information of each B cell informs the generation of human mAbs