Figure 5

Preparation and activity analysis of single-domain antibody micelles. (A) SDS-PAGE analysis of NCL of 626 μM Cys-PEG-DSPE with 50 μM sdAb-aGST-MESNA. The reaction was performed overnight at room temperature in buffer containing 100 mM sodium phosphate, 50 mM MPAA, 10 mM TCEP, pH 6. (B) Solid-phase binding assay of sdAb-aGST functionalized micelles to GST (squares). Micelle binding was monitored by measuring the fluorescence of the rhodamine lipids at 620 nm using an excitation of 578 nm. Control experiments using non-modified micelles incubated on GST (triangles) and sdAb-aGST functionalized micelles on milk powder coated plates (diamonds) are also shown for comparison. The solid line represents a fit to a 1:1 binding model using an apparent K _d of 14 μM.