Fabrication of self-assembled protein A monolayer and its application as an immunosensor.
The self-assembled layer of modified protein A was fabricated. In order to modify protein A, the surface group of protein A was substituted with thiol (-SH) functionality by using N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP) and dithiothreitol (DTT). The formation of a self-assembled protein A layer on a Au substrate and its increased binding capacity to antibody were confirmed by surface plasmon resonance (SPR) spectroscopy. The surface structure of self-assembled protein A layer, and the binding status of anti-bovine serum albumin (anti-BSA) and BSA were determined by atomic force microscopy (AFM). Treatment on the self-assembled protein A layer with a detergent, such as Tween 20, increased the binding capacity of anti-BSA, because protein A aggregation was reduced significantly by the detergent; this was confirmed by SPR spectroscopy. The self-assembled layer of chemically modified protein A with enhanced binding capacity can be used for immunosensor fabrication.
Lee
Woochang
W
Department of Chemical Engineering, Sogang University, C.P.O. Box 1142, Seoul 100-611, Republic of Korea.
Oh
Byung-Keun
BK
Min Bae
Young
Y
Paek
Se-Hwan
SH
Hong Lee
Won
W
Choi
Jeong-Woo
JW
D000906:Antibodies / Q000032:analysis* / Q000276:immunology; D000936:Antigen-Antibody Complex / Q000032:analysis / Q000276:immunology; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D020099:Coated Materials, Biocompatible / Q000138:chemical synthesis / Q000737:chemistry; D003460:Crystallization / Q000379:methods*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007118:Immunoassay / Q000295:instrumentation* / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000032:analysis / Q000276:immunology; D013205:Staphylococcal Protein A / Q000032:analysis* / Q000737:chemistry* / Q000276:immunology / Q000648:ultrastructure; D020349:Surface Plasmon Resonance / Q000379:methods*; D013499:Surface Properties
D000906:Antibodies; D000936:Antigen-Antibody Complex; D020099:Coated Materials, Biocompatible; D013205:Staphylococcal Protein A; D012710:Serum Albumin, Bovine