title: Magnetic gold nanoparticles in SERS-based sandwich immunoassay for antigen detection by well oriented antibodies.
abstract: The aim of the study was to develop an indirect, robust and simple in application method for the detection of bovine leukemia virus antigen gp51. Surface-enhanced Raman scattering (SERS) was applied as detection method. Magnetic gold nanoparticles (MNP-Au) modified by antibodies in oriented or random manner were used for the binding of gp51. The high performance liquid chromatography analysis indicated that the best antibody immobilization and antigen capturing efficiency was achieved using fragmented antibodies obtained after reduction of intact antibodies with dithiothreitol. In order to increase efficiency and sensitivity of immunoassay Raman labels consisting of gold nanorods coated by 5-thio-nitrobenzoic acid layer with covalently bounded antibodies have been constructed. The LOD and LOQ of the proposed immunoassay for antigen gp51 detection were found to be 0.95μgmL(-1) and 3.14μgmL(-1), respectively. This immunoassay was successfully applied for the detection of gp51 in milk samples in a rapid, reliable and selective manner. We believe that the proposed SERS-based immunoassay format can be applied for the detection of other proteins.
authors: NanoTechnas - Center of Nanotechnology and Materials Science, Faculty of Chemistry, Vilnius University, Naugarduko 24, LT-03225, Vilnius, Lithuania.
authors: Julija
authors: J
authors: Baniukevic
authors: Ismail
authors: I
authors: Hakki Boyaci
authors: Akif
authors: A
authors: Goktug Bozkurt
authors: Ugur
authors: U
authors: Tamer
authors: Arunas
authors: A
authors: Ramanavicius
authors: Almira
authors: A
authors: Ramanaviciene
mesh_terms: D000327:Adsorption; D000906:Antibodies / Q000737:chemistry*; D015374:Biosensing Techniques / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006046:Gold / Q000737:chemistry*; D007118:Immunoassay / Q000295:instrumentation*; D060526:Magnetic Fields; D053768:Metal Nanoparticles / Q000737:chemistry*; D013059:Spectrum Analysis, Raman / Q000295:instrumentation*
chemical_list: D000906:Antibodies; D006046:Gold