New immobilization method for immunoaffinity biosensors by using thiolated proteins

J. C. Pyun, S. D. Kim, J. W. Chung

Research output: Contribution to journalArticle

38 Citations (Scopus)

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 languageEnglish
Pages (from-to)227-233
Number of pages7
JournalAnalytical Biochemistry
Volume347
Issue number2
DOIs
Publication statusPublished - 2005 Dec 15

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Biosensing Techniques
Biosensors
Immobilization
Horseradish Peroxidase
Antibodies
Proteins
Antigens
Molecular recognition
Surface plasmon resonance
Biomolecules
Gold
Surface Plasmon Resonance
Charge transfer
Transducers
Assays
Binding Sites
Acids

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

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New immobilization method for immunoaffinity biosensors by using thiolated proteins. / Pyun, J. C.; Kim, S. D.; Chung, J. W.

In: Analytical Biochemistry, Vol. 347, No. 2, 15.12.2005, p. 227-233.

Research output: Contribution to journalArticle

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