Automated Antibody Isolation Using Magnetic Beads Attached Recombinant Human Protein(rhProtein) from Phage Display Libraries

摘要

The invariable yet increasing need for biological research, diagnostics, and therapeutic antibody development has been a drive to develop technologies such as hybridoma, phage display and transgenic mouse. Of these developments, phage display technology expresses functional antibody fragment (scFv, Fab) on the surface of filamentous phage, and its resulting phage library pool is used to screen for target specific antibodies. This process is called ‘biopanning’. Through this technology, a number of target specific antibodies have been discovered. Recently, automation and several high-throughput method have been applied to antibody phage display technology to improve its speed and efficiency. We performed bio-panning based on automated system using recombinant human protein(Target X) attached to magnetic beads through a pin-based magnetic particle processor. We used two synthetic scFv phage libraries to perform four rounds of bio-panning each. After the automated bio-panning, we screened 752 scFvs against target X using affinity ELISA assay, which isolated 19 different target X specific clones. In contrast, generally used biopanning method with immobilized antigen onto immunotube did not isolate any target X specific scFvs. Through this study, we could isolate the target X specific scFv which has not been found through the bio-panning method with immobilized antigen with the automated bio-panning. This automated bio-panning procedure improves the throughput of target specific scFv candidates and thus allows the selection of scFvs against many different targets in parallel with high efficiency.