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. Author manuscript; available in PMC: 2019 Aug 28.
Published in final edited form as: ACS Nano. 2018 Jan 26;12(2):1544–1563. doi: 10.1021/acsnano.7b08122

Figure 5.

Figure 5.

In vitro evaluation of the α-CD20(DBCO)10 and PAMAM(D-89Y)8(N)29(Rhod)8-based pretargeted system. (a) Flow cytometry histograms of α-CD20(DBCO)10-pretreated Raji cells after incubation with different concentrations of (i) azide-functionalized A488 and (ii) PAMAM(D-89Y)8(N)29(Rhod)8 containing 11.6–23,220 nM of terminal azide. (iii) The plot of MFIs of α-CD20(DBCO)10-pretreated Raji cells after being incubated with different concentrations of azide-functionalized A488 and PAMAM(D-89Y)8(N)29(Rhod)8, as determined by the flow cytometry method. (b) Representative CLSM images of (i) unstained Raji cells (control); (ii) Raji cells stained with α-CD20(DBCO)10(A488)2 for 1 h, washed, with and without further incubation at 37 °C for another 6 h; (iii) Raji cells pretreated with α-CD20(DBCO)10(A488)2 for 1 h before staining with PAMAM(D-89Y)8(N)29(Rhod)8 for another 1 or 6 h; (iv) Raji cells incubated with the antibody-dendrimer premixture for 1 or 6 h; and (v) Raji cells incubated with preblocked α-CD20(DBCO)10(A488)2 for 1 h before being incubated with PAMAM(D-89Y)8(N)29(Rhod)8 for another 1 h.