PRODUCTION OF MONOCLONAL RECOMBINANT ANTIBODIES WITHOUT THE USE OF HYBRIDOMAS BY $i(IN VITRO) SPLEEN FRAGMENT CULTURE COMBINED WITH ISOTHERMAL SELF-SUSTAINED SEQUENCE REPLICATION OF RNA

摘要

The invention relates to a method for producing a recombinant protein having a preselected specificity and binding affinity with respect to a given antigen. The method includes generally combine culturing a spleen fragment, for the in vitro somatic mutation of B cells in response to antigen, and the preferential amplification of RNA encoding heavy and light chains of antibodies required produced by these cells. The amplified RNA is converted into DNA and then incorporated into expression vectors; transfected host cells then express chains of single-chain antibody or an antibody having the preset characteristics. The invention also relates to groups of generic degenerate primers for the amplification of RNA encoding the mouse IgG.