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[Preprint]. 2022 Feb 1:2021.06.28.450181. [Version 4] doi: 10.1101/2021.06.28.450181

Figure 2.

Figure 2.

A) Assessment of efficiency of SpyTag/SpyCatcher coupling of RBD onto HbpD of OMVs. RBD-Spy-His and His-Spy-RBD were coupled to Hbp-SpyCatcher OMVs. Proteins of conjugated and non-conjugated OMVs were separated by SDS-PAGE and stained with Coomassie Brilliant Blue. RBD-HbpD appears as a ~160 kDa band, while free HbpD is seen as a ~125 kDa band. Densitometry suggested that approximately 90% or more of HbpD was coupled with RBD in the conjugated populations compared with unconjugated OMVs (rightmost lane). Other outer membrane proteins of OMVs (OMPs) are indicated; B) Coomassie Brilliant Blue staining of SDS-PAGE gel containing non-conjugated OMVs and a 1:1 mixture of RBD-Spy-His and His-Spy-RBD-coupled OMVs.