Abstract
Abstract Human monoclonal antibodies have great potential for use in the treatment of various human diseases. An in vitro immunisation protocol using human peripheral blood mononuclear cells (PBMCs) was developed to generate human antigen‐specific antibodies. After antigen sensitisation of PBMCs in vitro , B cells producing antigen‐specific antibody can be propagated within a week. Heavy and light chain variable region genes were easily amplified from these in vitro immunised PBMCs. After generating a combinatorial phage library, antigen‐specific clones were selected by panning rounds. Next, the variable region genes of heavy and light chains of these selected clones were combined with human IgG constant region genes to produce human IgG‐type antibody in mammalian cells. This in vitro immunisation method will be applied widely for the production of human monoclonal antibodies for therapeutics by combination with several display methods. Key Concepts: In vitro immunisation method can be used to induce B cells producing antigen‐specific human monoclonal antibodies from human peripheral lymphocytes. Phage display method can be used to efficiently generate antigen‐specific human monoclonal antibodies. The combined use of in vitro immunisation and phage display methods is beneficial to efficiently establish recombinant animal cells producing antigen‐specific human monoclonal antibodies from a relatively small library.
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