title: Dextran as a generally applicable multivalent scaffold for improving immunoglobulin-binding affinities of peptide and peptidomimetic ligands.
abstract: Molecules able to bind the antigen-binding sites of antibodies are of interest in medicine and immunology. Since most antibodies are bivalent, higher affinity recognition can be achieved through avidity effects in which a construct containing two or more copies of the ligand engages both arms of the immunoglobulin simultaneously. This can be achieved routinely by immobilizing antibody ligands at high density on solid surfaces, such as ELISA plates, but there is surprisingly little literature on scaffolds that routinely support bivalent binding of antibody ligands in solution, particularly for the important case of human IgG antibodies. Here we show that the simple strategy of linking two antigens with a polyethylene glycol (PEG) spacer long enough to span the two arms of an antibody results in higher affinity binding in some, but not all, cases. However, we found that the creation of multimeric constructs in which several antibody ligands are displayed on a dextran polymer reliably provides much higher affinity binding than is observed with the monomer in all cases tested. Since these dextran conjugates are simple to construct, they provide a general and convenient strategy to transform modest affinity antibody ligands into high affinity probes. An additional advantage is that the antibody ligands occupy only a small number of the reactive sites on the dextran, so that molecular cargo can be attached easily, creating molecules capable of delivering this cargo to cells displaying antigen-specific receptors.
authors: Departments of Chemistry and Cancer Biology, The Scripps Research Institute , 130 Scripps Way, Jupiter, Florida 33458, United States.
authors: Jumpei
authors: J
authors: Morimoto
authors: Mohosin
authors: M
authors: Sarkar
authors: Sophia
authors: S
authors: Kenrick
authors: Thomas
authors: T
authors: Kodadek
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002999:Clone Cells; D003911:Dextrans / Q000737:chemistry*; D019281:Dimerization; D057809:HEK293 Cells; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology*; D007945:Leukemia, Lymphoid / Q000473:pathology; D008024:Ligands; D051379:Mice; D008958:Models, Molecular; D010455:Peptides / Q000737:chemistry* / Q000276:immunology*; D057786:Peptidomimetics / Q000737:chemistry* / Q000276:immunology*; D011092:Polyethylene Glycols / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation
chemical_list: D003911:Dextrans; D007074:Immunoglobulin G; D008024:Ligands; D010455:Peptides; D057786:Peptidomimetics; D011092:Polyethylene Glycols