A polymer surface for antibody detection by using surface plasmon resonance via immobilized antigen

• Published in: Current applied physics 2013, 13(6), , pp.1008-1013
• Main Authors: Puttharugsa, Chokchai, Wangkam, Thidarat, Houngkamhang, Nongluck, Yodmongkol, Sirisa, Gajanandana, Oraprapai, Himananto, Orawan, Sutapun, Boonsong, Amarit, Ratthasart, Somboonkaew, Armote, Srikhirin, Toemsak
• Format: Article
• Language: English
• Subjects: Acidovorax avenae subsp. citrulli; Acrylic acid; Antibodies; Antibody detection; antibody formation; Antigens; bacteria; Binding; bovine serum albumin; Density; detection limit; image analysis; Imaging; immobilized antigens; monoclonal antibodies; Optical biosensors; Physical adsorption; physics; Plasmons; polymers; Screening; surface plasmon resonance; Surface plasmon resonance (SPR); 물리학
• ISSN: 1567-1739; 1878-1675
• DOI: 10.1016/j.cap.2013.02.003

A polymer substrate based surface plasmon resonance (SPR) technique was developed for detection of specific monoclonal antibody 10B2 (MAb 10B2) against bacterium Acidovorax avenae subsp. citrulli (Aac). The monolayer of Aac antigen was physically immobilized on 95:5 polystryrene – copoly acrylic acid (95PSMA) for detection of antibody. The amount of antigen–antibody binding was found to depend on the surface density of immobilized Aac on the sensor surface and the antibody concentration. The detection limit was 5 μg/ml which was lower than the required concentration during the normal production of the antibody at 10–100 μg/ml. This suggests a possible use of surface for the antibody screening. Moreover, an application in antibody screening was explored by combination of surface plasmon resonance imaging (SPR imaging) and antibody detection assay on the 95PSMA surface. Two antigens of bovine serum albumin (BSA) and Aac were used as a model system for antibody screening. The result shows that both antibodies can be distinguished using the immobilized antigens on the 95PSMA surface based SPR imaging technique. ► Antibody was detected by SPR technique via an immobilized antigen on polymer surface. ► The signal of antibody detection depended on the immobilized antigen density surface. ► Application in antibody screening was explored by using SPR imaging.