Enhancement of the protein loading density by a pre-cleaning process of a gold substrate: confocal laser scanning microscopic study.
The immobilization of glucose oxidase (GOx), using self assembled monolayers (SAMs) on gold surfaces, was investigated by grazing angle FT-IR spectroscopy, surface plasmon resonance (SPR) spectroscopy, and atomic force microscopy (AFM) in conjunction with confocal laser scanning microscopy (CLSM). To find an optimum condition for the maximum GOx loading density on gold surfaces, different cleaning protocols were examined. The loading density of GOx on surfaces was investigated by AFM and CLSM. In particular, CLSM was more effective for identifying the GOx density than AFM, since its scanning speed is much faster and it covers a larger area. Based on CLSM images of the GOx immobilized on the surfaces, it was concluded that the pre-cleaning process of gold substrates using different solvents, such as acetone, ethanol and 2-propanol, is very important for enhancing the GOx loading density. This result enables us to investigate an effective fabrication process in fabricating biosensors.
Department of Applied Chemistry, Hanyang University, Ansan 426-791, South Korea.
Taehan
T
Park
Jaebum
J
Choo
Moonkwon
M
Lee
Yang S
YS
Kim
Eun Kyu
EK
Lee
Hae Seong
HS
Lee
D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D004800:Enzymes, Immobilized / Q000737:chemistry*; D005949:Glucose Oxidase / Q000737:chemistry*; D006046:Gold; D018625:Microscopy, Atomic Force; D018613:Microscopy, Confocal; D017550:Spectroscopy, Fourier Transform Infrared; D020349:Surface Plasmon Resonance; D013499:Surface Properties
D004800:Enzymes, Immobilized; D006046:Gold; D005949:Glucose Oxidase