Immunized Phage Display

Custom immunized phage display antibody libraries are prepared by combining the immunological and the phage display techniques together. With this, the animal is immunized with the antigen of interest. After testing the immunogenic response  by ELISA and or western blotting,  PMBC or the spleen is isolated and the cDNA corresponding to vL and vHgenes are amplified using a set of specific primers and cloned into a phagemid vector.  The inserted cDNA is modified to facilitate the expression and purification of antibodies from bacterial or human cell lines.  The libraries prepared in this way are of moderate size and populated with the clones expressing high affinity antibodies on the surface of the phages which could be quickly screened. The sequence of the antibodies isolated by this method can be easily modified, humanized or chimerized.  The appropriate clone can be used for large scale production.