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. Author manuscript; available in PMC: 2016 Nov 23.
Published in final edited form as: Chemistry. 2015 Oct 16;21(48):17178–17183. doi: 10.1002/chem.201503057

Figure 2.

Figure 2

Uniform attachment and characterization of a murine Class II MHC-specific targeting nanobody, VHH7, on GO nanosheets. a, Overview of sortase-mediated site-specific ligation of VHH7 onto GO nanosheets. VHH7 was C-terminally modified via a sortase-mediated ligation to install a TAMRA fluorophore and an azide. The GO nanosheet was functionalized with a DBCO handle for a “click” reaction with the azide-modified VHH7. b, AFM images of GO nanosheets and GO nanosheets with VHH7 immobilized on silicon substrates. The height difference between GO nanosheets with or without VHH7 immobilization was ~20–80 nm. The topography of the slide observed under the AFM is characteristic of protein bound to GO.[6] c, Whole blood analysis from Class II MHC-eGFP knock-in mice. A small volume of blood (30 μL) was loaded into the assembled chamber and incubated at 37°C for 10 min. The chamber was then disassembled and the cell-bearing substrate slide was gently washed 3x with PBS. After washing, cells were analyzed by fluorescence microscopy and quantitative image analysis.