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The
utilization of BSA-modified chip on the investigation of
ligand/protein interaction with surface plasma resonance
LiHua Chen,
Qiang Wang and WanGuo Hou*
College
of Chemistry and Molecular Engineering, Qingdao University
of Science and Technology, Qingdao, Shandong, People’s
Republic of China.
*Corresponding author. E-mail:
lihuachen@qust.edu.cn.
Tel: +86 15054246089. Fax: +86 532 84023927.
Abbreviations: GSH,
glutathione; GST, glutathione-S-trans- ferase; SPR,
surface plasmon resonance; NSB, nonspecific
binding; BSA, bovine serum albumin;
SAMs,
self-assembled monolayers;
PDEA,
2-(2-pyridinyldithio) ethane amine hydro- chloride; BS3,
Bis(3-sulfo-N-hydroxysuccinimide ester) sodium salt;
GMBS, suberic acid 4-maleimidobutyric acid N-hydro-
xysuccinimide ester; PBS, phosphate buffered saline
tablets; DMSO, dimethylsulfoxide; EDC,
1-ethyl-3-[3- dimethylamino-
propyl]carbodiimide hydrochloride;
NHS,
N-hydroxysuccini- mide.
Accepted 20 November, 2009 |
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The kinetic behavior of glutathione (GSH)/ glutathione-S-transferase
(GST) was investigated using surface plasmon resonance (SPR).
Here, an alkanethiol-modified chip incorporated with bovine
serum albumin (BSA) was employed. Subsequently, GSH was
anchored on BSA surface only in the experimental channel and
the without-active BSA surface was designed as the reference
channel to improve the quality of the binding data and
prevent a number of experimental artifacts to complicate the
final biosensor analysis. Our results demonstrated that the
BSA-modified chip was effective not only in binding the
target proteins but also in suppressing the nonspecific
binding (NSB) of proteins.
Key words:
Surface plasmon resonance, bovine serum albumin,
glutathione, glutathione-S-transferase, alkanethiol. |
