Figure 4. Specificity of antigenic selection during mRNA/cDNA display.

The mutant library that was constructed based on the Nb-GFP template was displayed using the mRNA/cDNA display method described in this manuscript, and exposed to either GFP or mouse P-glycoprotein (mP-gp), which is an integral membrane transporter protein purified as previously published¹⁸. Following washes and elution, the binder sequences were regenerated using PCR. The agarose gel shows that for the same input material (library and RT reaction), Nb-specific binders were obtained for selection with GFP as the antigen, 40% sequences of which were confirmed to be Wt Nb-GFP, whereas Nb-specific PCR product was absent following exposure to mP-gp as the antigen, confirming the mRNA/cDNA display method was successful in removing non-specific binder Nbs.