Protein attach to nanodiamonds as pseudo stationary phase for neuro-disease diagnosis.

ROMINA LÓPEZ; LORENA L. SOMBRA; PATRICIA W. STEGE

Cartagena

Simposio; 21st Latin American Symposium on Biotechnology, Biomedical, Biopharmaceutical and Industrial Applications of Capillary Electrophoresis and Microchip Technology; 2015

In the last two decades, significantinterest has been reported on the use of nanometer sized diamond particles(NDs). These nanoparticles have attracted the interest of the nanoscience andnanotechnology communities, because of the potential usein the manufacturing ofbright, low voltage (cold) cathodes and light emitters nanodiamond films. Furthermore,chemical modifications of the surface of the nanoparticlesare becomingofsignificant value for the syntesis of bionanoconjugates.Glucose transporter type 1(Glut1) deficiency syndrome is a rare genetic metabolic disorder characterized bydeficiency of a protein that is required for glucose to cross the blood-brain barrier.In children affects the nervous system and can be manifested with a variety ofneurological symptoms.  The most common symptomis seizures, which usually begin within the first few months of life. However,the symptoms and severity of Glut1 deficiency syndrome can vary substantially fromone person to another. If diagnoses in the early stage of the disease formation,it can be of help to improve the quality of life of patients.The aim of this study was to apply NDs as animmunosupport forthe determination of Glut1 in a red cells lysate using an affinitycapillary electrochromatography technique.The Glut1 antibodies were attached tothe NDs and the bionanoconjugate was added to the background electrolyte (BGE)asa pseudostationary phase (PSP).A Beckman P/ACE MDQ capillaryelectrophoresis instrument was used.The dimensions of the capillary were50 µm i.d. and15 cm total lenght, and the PSP system consisted of sodium phosphate 40 mM, pH=7.4, and 7%NDs-Glut1antibodies covalently bonded by the carboxylated groups present in thenanodiamonds and the amine groups present in the antibodies. The proposed methodwas successfully employed for the detection of normal Glut1 in a red cellslysate. The use of NDs-Glut1antibodies as pseudo stationary phases in nanoelectrokinetic(nanoEKC) allows for the determination of the protein of interest with minimaltreatment of the samples.