Evaluation of different strategies for magnetic particle functionalization with DNA aptamers.
The optimal bio-functionalization of magnetic particles is essential for developing magnetic particle-based bioassays. Whereas functionalization with antibodies is generally well established, immobilization of DNA probes, such as aptamers, is not yet fully explored. In this work, four different types of commercially available magnetic particles, coated with streptavidin, maleimide or carboxyl groups, were evaluated for their surface coverage with aptamer bioreceptors, efficiency in capturing target protein and non-specific protein adsorption on their surface. A recently developed aptamer against the peanut allergen, Ara h 1 protein, was used as a model system. Conjugation of biotinylated Ara h 1 aptamer to the streptavidin particles led to the highest surface coverage, whereas the coverage of maleimide particles was 25% lower. Carboxylated particles appeared to be inadequate for DNA functionalization. Streptavidin particles also showed the greatest target capturing efficiency, comparable to the one of particles functionalized with anti-Ara h 1 antibody. The performance of streptavidin particles was additionally tested in a sandwich assay with the aptamer as a capture receptor on the particle surface. While the limit of detection obtained was comparable to the same assay system with antibody as capture receptor, it was superior to previously reported values using the same aptamer in similar assay schemes with different detection platforms. These results point to the promising application of the Ara h 1 aptamer-functionalized particles in bioassay development.
Department of Biosystems-MeBioS-Biosensor Group, KU Leuven, Leuven, Belgium.
Elena
E
Pérez-Ruiz
Department of Biosystems-MeBioS-Biosensor Group, KU Leuven, Leuven, Belgium. Electronic address: jeroen.lammertyn@biw.kuleuven.be.
Jeroen
J
Lammertyn
Department of Biosystems-MeBioS-Biosensor Group, KU Leuven, Leuven, Belgium.
Dragana
D
Spasic
D000485:Allergens / Q000032:analysis / Q000235:genetics; D052179:Antigens, Plant / Q000032:analysis / Q000235:genetics; D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000235:genetics; D001681:Biological Assay / Q000379:methods; D015374:Biosensing Techniques / Q000379:methods*; D001709:Biotechnology; D006023:Glycoproteins / Q000032:analysis / Q000235:genetics; D006801:Humans; D008280:Magnetics; D008565:Membrane Proteins; D021183:Peanut Hypersensitivity / Q000276:immunology; D010940:Plant Proteins / Q000032:analysis / Q000235:genetics
D000485:Allergens; D052179:Antigens, Plant; D052157:Aptamers, Nucleotide; C500889:Ara h 1 protein, Arachis hypogaea; D006023:Glycoproteins; D008565:Membrane Proteins; D010940:Plant Proteins
Aptamer; Biofunctionalization; DNA; Magnetic particles