Exp Mol Med.
1999 Sep;31(3):122-125.
A radioimmunoassay method for detection of DNA based on chemical immobilization
of anti-DNA antibody
- Affiliations
-
- 1Department of Chemistry, Kyung Hee University, Seoul, Korea.
Abstract
-
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.