title: Chemoselective surface immobilization of proteins through a cleavable peptide.
abstract: Surface immobilization of biomolecules is a fundamental step in several experimental techniques such as surface plasmon resonance analysis and microarrays. Oxime ligation allows reaching chemoselective protein immobilization with the retention of native-like conformation by proteins. Beside the need for chemoselective ligation of molecules to surface/particle, equally important is the controlled release of the immobilized molecules, even after a specific binding event. For this purpose, we have designed and assessed in an SPR experiment a peptide linker able to (i) anchor a given protein (enzymes, receptors, or antibodies) to a surface in a precise orientation and (ii) release the immobilized protein after selective enzymatic cleavage. These results open up the possibility to anchor to a surface a protein probe leaving bioactive sites free for interaction with substrates, ligands, antigens, or drugs and successively remove the probe-ligand complex by enzymatic cleavage. This peptide linker can be considered both an improvement of SPR analysis for macromolecular interaction and a novel strategy for drug delivery and biomaterial developments.
authors.affiliation: Department of Chemical Process Engineering, University of Padova, Via Marzolo 9, Padua, Italy. monica.dettin@unipd.it
authors.forename: Monica
authors.initials: M
authors.lastname: Dettin
authors.forename: Natali
authors.initials: N
authors.lastname: Muncan
authors.forename: Antonella
authors.initials: A
authors.lastname: Bugatti
authors.forename: Francesca
authors.initials: F
authors.lastname: Grezzo
authors.forename: Roberta
authors.initials: R
authors.lastname: Danesin
authors.forename: Marco
authors.initials: M
authors.lastname: Rusnati
mesh_terms: D000941:Antigens / Q000737:chemistry; D054377:Chemokine CXCL12 / Q000737:chemistry; D006046:Gold; D055767:Immobilized Proteins / Q000737:chemistry*; D008024:Ligands; D020778:Matrix Metalloproteinase 2 / Q000737:chemistry; D010455:Peptides / Q000737:chemistry*; D011732:Pyridoxal Phosphate / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry; D060327:Solid-Phase Synthesis Techniques; D020349:Surface Plasmon Resonance
chemical_list: D000941:Antigens; D054377:Chemokine CXCL12; D055767:Immobilized Proteins; D008024:Ligands; D010455:Peptides; D012710:Serum Albumin, Bovine; D011732:Pyridoxal Phosphate; D006046:Gold; D020778:Matrix Metalloproteinase 2