Figure 2.

Construction and application of cytosolic/nuclear camelid nanobodies. Until now, most cytosolic camelid nanobodies have been constructed after immunization of camels. The VHH domains will be amplified from the cDNA isolated from the peripheral blood lymphocytes of immunized camels. Then a phage display repertoire is generated to display the VHH repertoire on the surface. Biopanning with the antigen will deliver specific recombinant nanobody phages from which the genes can be cloned into an appropriate targeting vector for expression in the cytosol or nucleus. The function of the intrabody can be tested in cell culture or applied in a xenograft tumor mouse model. Alternatively, a transgenic mouse expressing the nanobody can be generated. (P, promoter; R, resistant gene; Nb, nanobody; NLS, nucleus leader sequence).