Aryl diazonium for biomolecules immobilization onto SPRi chips.
A method for the immobilization of proteins at the surface of surface plasmon resonance imaging (SPRi) chips is presented. The technology, based on the electro-deposition of a 4-carboxymethyl aryl diazonium (CMA) monolayer is compared to a classical thioctic acid self-assembled monolayer. SPRi live recording experiments followed by the quantification of the diazonium surface coverage demonstrate the presence of a monolayer of electro-deposited molecules (11*10(12) molecules mm(-2)). This monolayer, when activated through a classical carbodiimide route, generates a surface suitable for the protein immobilization. In the present study, protein A and BSA are immobilized as specific and control spots (150 microm id), respectively. The AFM characterization of the spots deposited onto CMA or thioctic acid modified chips prove the presence of 4.7 nm protein monolayers. Finally, the SPRi detection capabilities of the two surface chemistries are compared according to specific signal, non-specific interaction and regeneration possibilities. Advantages are given to the CMA surface modification since no measurable non-specific signal is obtained while reaching a higher specific signal.
Mandon
Céline A
CA
Institut de Chimie et Biochimie Moléculaires et Supramoléculaires CNRS 5246 ICBMS, Université Lyon 1, Bâtiment CPE 43, bd du 11 novembre 1918-69622 Villeurbanne, France.
Blum
Loïc J
LJ
Marquette
Christophe A
CA
D000818:Animals; D002417:Cattle; D003979:Diazonium Compounds / Q000737:chemistry*; D055767:Immobilized Proteins / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D013205:Staphylococcal Protein A / Q000737:chemistry*; D020349:Surface Plasmon Resonance
D003979:Diazonium Compounds; D055767:Immobilized Proteins; D013205:Staphylococcal Protein A; D012710:Serum Albumin, Bovine