Regioselective covalent immobilization of recombinant antibody-binding proteins A, G, and L for construction of antibody arrays.

摘要: Immobilized antibodies are useful for the detection of antigens in highly sensitive microarray diagnostic applications. Arrays with attached regioselectively a uniform orientation typically more than those random orientations. Direct regioselective immobilization on solid support requires modified form protein. We now report general approach attachment to surface using truncated forms antibody-binding proteins A, G, and L that retain structural motifs required antibody binding. The recombinant have C-terminal CVIX protein farnesyltransferase recognition motif allows us append bioorthogonal azide or alkyne moiety use Cu(I)-catalyzed Huisgen cycloaddition attach binding suitably glass surface. This offers several advantages. produced Escherichia coli, chemoselectively mo...