MEASUREMENT OF ENZYMATIC ACTIVITY USING GOLD NANOPARTICLES IN OPEN TUBE CAPILLARY ELECTROPHORESIS

FELICI, EMILIANO; CHIEN CHUN WANG; GOMEZ, MARÍA ROXANA; STEGE, PATRICIA

Simposio; 21th Latin-American Symposium on Biomedical, Biopharmaceutical and Industrial Applications of Capillary Electrophoresis (LACE); 2015

Nanostructured materials have great interest in scientific-technological field due to their physical properties. Metal based nanoparticles, especially gold nanoparticles (AuNPs) have excellent biocompatibility with different biological molecules. Our particular interest in this type of nanoparticles relies on the possibility of modify their surfaces with the aim of acting as support of different macromolecules bringing stability to the system.The aim of this work was the immobilization of Glutathione reductase-AuNPs bionanocojugated AuNPs to the inner phase of the 50 μm i.d. and 15 cm total lenght capillary using layer by layer polymeric network in the inlet side. AuNPs were synthesized by reduction with sodium citrate added under stirring with excess of NaBH4. The AuNPs were purified and the solution obtained was characterized by UV-Vis spectrophotometry and capillary electrophoresis. Bionanocojugated was attached in the inner wall of capillaries using dimethylsiloxane as polymeric agent. Layer-by-Layer (LbL) deposition was developed by immersion procedure of the inlet side of the capillary. The immobilized Bionanoconjugate was characterized by SEM to contact angle measurement observing morphological features designed biosensor.AuNPs have the advantage of being stable, having low cost and high biocompatibility acting as support and stabilizing biological molecules increasing enzyme activity. In addition, the matrix used in the immobilization process provides a biocompatible environment which allows the enzyme remain active after the immobilization and improving the sensitivity of the biosensor