Abstract
A new immobilization method for immunoaffinity (IA) biosensors that ensures the high surface density and the stability of the IA layer was developed. For the immobilization of biomolecules, the molecular recognition protein was first thiolated by covalent conjugation of mercaptopropionic acid, and then the thiolated protein was attached on the gold surface of the transducer. In this work, horseradish peroxidase (HRP) and its antibody were used as a model antigen-antibody, and the following properties of the IA layer prepared by thiolated protein were estimated: (i) biological integrity of HRP after the immobilization process by using activity assay, (ii) charge transfer resistance by immobilization, (iii) mass loading by the surface plasmon resonance (SPR) biosensor, (iv) number of binding sites, and (v) feasibility test for the measurement of capacitive change by the antigen-antibody interaction. Based on these parameters, the immobilization method by using thiolated protein was determined to be feasible for application to IA biosensors.
| Original language | English |
|---|---|
| Pages (from-to) | 227-233 |
| Number of pages | 7 |
| Journal | Analytical Biochemistry |
| Volume | 347 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - 2005 Dec 15 |
Bibliographical note
Funding Information:The authors are thankful for the financial support of the Ministry of Science and Technology (MOST), Korea, and the Korean Institute of Science and Technology (KIST), Korea.
All Science Journal Classification (ASJC) codes
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology