Abstract
The reversible, oriented immobilization of proteins on solid surfaces is a prerequisite for the investigation of molecular interactions and the controlled formation of supramolecular assemblies. This paper describes a generally applicable method using a synthetic chelator thioalkane that can be self-assembled on gold surfaces. The reversible binding of an anti-lysozyme F(ab) fragment modified with a C-terminal hexahistidine extension was monitored and the apparent dissociation constants determined using surface plasmon resonance. Infra-red spectroscopy demonstrated that the secondary structure of the protein was unaffected by the immobilization process. The retention of functionality of the immobilized protein was also successfully demonstrated. Given the mild reaction conditions and reversibility, this method of oriented immobilization of proteins opens possibilities for the development of biosensors.
Original language | English |
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Pages (from-to) | 155-161 |
Number of pages | 7 |
Journal | Biosensors and Bioelectronics |
Volume | 14 |
Issue number | 2 |
DOIs | |
State | Published - Feb 1999 |
Externally published | Yes |
Keywords
- ATR
- Attenuated total reflection
- CTA
- Chelator thioalkane
- FTIR
- Fourier transform infra-red spectroscopy
- HTA
- His-Tag
- Histidine- tag
- Hydroxythioalkane
- NTA
- Nitrilotriacetic acid
- RU
- Resonance unit
- SA
- SAM
- SPR
- Self- assembled monolayer
- Self-assembly
- Surface plasmon resonance
- TFA
- Trifluoroacetic acid