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. Author manuscript; available in PMC: 2016 Jul 1.
Published in final edited form as: Biochim Biophys Acta. 2015 Mar 26;1850(7):1397–1404. doi: 10.1016/j.bbagen.2015.03.009

Figure 3. Analysis of the extent of the in vivo biotinylation of VHH T7.

Figure 3

T7bt: T7 biotinylated in vivo and purified on Ni-NTA-agarose. T7100%bt: fraction of T7bt eluted from monomeric avidin-agarose. T7−bt: VHH T7 expressed without addition of biotin and purified on Ni-NTA-agarose. These three preparation of VHH T7 were titrated against TCC-BSA by ELISA using streptavidin-HRP for developing; T7bt, circles; T7100%bt, squares; T7−bt, triangles. The insert shows the SDS-PAGE analysis of the different preparations of VHH T7 applied to avidin-agarose spin columns; Vd, void; Ad, bound fraction eluted with SDS-PAGE sample buffer. The upper (T7) and lower (Av) bands correspond to the VHH, and the avidin monomer that is released from the avidin-agarose.