A radioimmunoassay method for detection of DNA based on chemical immobilization of anti-DNA antibody.
High selectivity provided by biomolecules such as antibodies and enzymes has been exploited during the last two decades for development of biosensors. Of particular importance are efficient immobilization methods for biomolecules in order to preserve their biological activities. In this study, we have evaluated immobilization strategies for an anti-DNA antibody on a self-assembled monolayer of omega-functionalized thiols. The antibody was immobilized via peptide bond formation between the primary amines in the antibody and the carboxyl groups on the self-assembled monolayer. The peptide bond coupling was achieved by activating COOH groups on the surface through N-Hydroxysuccimide (NHS)-ester formation, followed by acylation of NH2 group in the antibody. DNA binding activity of the immobilized antibody was examined by counting beta emission from 35S-labeled DNA.
Yoo
S K
SK
Department of Chemistry, Kyung Hee University, Seoul, Korea.
Yoon
M
M
Park
U J
UJ
Han
H S
HS
Kim
J H
JH
Hwang
H J
HJ
D000974:Antibodies, Antinuclear; D004247:DNA / Q000032:analysis* / Q000276:immunology; D004268:DNA-Binding Proteins / Q000737:chemistry; D006046:Gold; D008567:Membranes, Artificial; D016133:Polymerase Chain Reaction; D011145:Polyvinyls / Q000737:chemistry; D011863:Radioimmunoassay / Q000379:methods*; D008063:Thioctic Acid / Q000737:chemistry; D005465:Fluorocarbon Polymers
D000974:Antibodies, Antinuclear; D004247:DNA; D004268:DNA-Binding Proteins; D006046:Gold; D008567:Membranes, Artificial; D011145:Polyvinyls; D008063:Thioctic Acid; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers