Magnetic gold nanoparticles in SERS-based sandwich immunoassay for antigen detection by well oriented antibodies


Baniukevic J., BOYACI İ. H., Bozkurt A. G., TAMER U., Ramanavicius A., Ramanaviciene A.

BIOSENSORS & BIOELECTRONICS, cilt.43, ss.281-288, 2013 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 43
  • Basım Tarihi: 2013
  • Doi Numarası: 10.1016/j.bios.2012.12.014
  • Dergi Adı: BIOSENSORS & BIOELECTRONICS
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.281-288
  • Anahtar Kelimeler: SERS, Immunoassay, Magnetic gold nanoparticles, Oriented immobilization, BLV, ENHANCED RAMAN-SCATTERING, BOVINE LEUKEMIA-VIRUS, INTERNAL-REFLECTION ELLIPSOMETRY, DAIRY-CATTLE, IMMUNOGOLD NANOPARTICLES, IMMOBILIZATION, FRAGMENTS, INFECTION, PROTEIN, ASSAY
  • Gazi Üniversitesi Adresli: Evet

Özet

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 0.95 mu g mL(-1) and 3.14 mu g mL(-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. (c) 2012 Elsevier B.V. All rights reserved.