Selected article for: "approach stabilize and binding activity"
Author: Chu, Ruiyin; Reczek, David; Brondyk, William
Title: Capture-stabilize approach for membrane protein SPR assays
  • Document date: 2014_12_8
    • ID: ueofl0wn_6
    Snippet: Capture-stabilized CD52 in virus-like particles. We further extended the application of the capture-stabilize approach to viruslike particles for analyzing other types of membrane protein targets in their native membrane environment. Human CD52, a heavily glycosylated glycosylphosphatidylinositol (GPI)-anchored protein composed of only 12 AA residues 26, 27 , was used as an example. As shown in Figure 5 , CD52 virus-like particles (VLPs) were pre.....
    Document: Capture-stabilized CD52 in virus-like particles. We further extended the application of the capture-stabilize approach to viruslike particles for analyzing other types of membrane protein targets in their native membrane environment. Human CD52, a heavily glycosylated glycosylphosphatidylinositol (GPI)-anchored protein composed of only 12 AA residues 26, 27 , was used as an example. As shown in Figure 5 , CD52 virus-like particles (VLPs) were prepared from 293FT cells. After Western blot analysis to verify the presence of CD52 in the VLP prep, VLPs containing CD52 were captured by the anti-CD52 antibody Alemtuzumab 26 on a C1 chip surface. The captured VLPs were then stabilized by the same approach. Three different anti-CD52 monoclonal antibodies with slightly different binding epitopes (Chu, et al., 2014, unpublished data), including the capturing antibody Alemtuzumab, demonstrated good binding activity in kinetic assays. Similar to Ni-affinity capturing of the purified hCXCR5, the capture of CD52 VLPs by Alemtuzumab is target specific. The post capture cross-linking step stabilizes CD52 in the VLP surface and enables multiple kinetic binding assay cycles to be performed.

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