Figure 5.

Isolation of specific internalizing antibodies by phage display. The antibody-phage library was initially subtracted on healthy cells, and then the free phage were screened on cells harbouring the antigen of interest. A washing step was performed to remove all the non-specific and unbound phage. All the previous steps were conducted at 4 °C. The cells were then incubated at 37 °C to induce phage internalization. Cell surface bound phage were removed by low pH elution buffer, while the internalized phage were retrieved by cell lysis. The isolated phage particles were then amplified and used for subsequent panning rounds. Figure shows methodology described by Fitting et al. [130].