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A polymer surface for antibody detection by using surface plasmon resonance via immobilized antigen |
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| Authors: | Chokchai Puttharugsa Thidarat Wangkam Nongluck Houngkamhang Sirisa Yodmongkol Oraprapai Gajanandana Orawan Himananto Boonsong Sutapun Ratthasart Amarit Armote Somboonkaew Toemsak Srikhirin |
| Institution: | 1. Center of Intelligent Materials and Systems, Nanotec Center of Excellence at Mahidol University, Bangkok, Thailand;2. Physics Department, Faculty of Science, Mahidol University, Rama VI Rd., Phayathai, Bangkok 10400, Thailand;3. Materials Science and Engineering Programme, Faculty of Science, Mahidol University, Rama VI Rd., Phayathai, Bangkok 10400, Thailand;4. Monoclonal Antibody Production Laboratory, National Center for Genetic Engineering and Biotechnology (BIOTEC), Pathumthani 12120, Thailand;5. School of Electronic Engineering and School of Telecommunications Engineering, Suranaree University of Technology, 111 University Avenue, Muang District, Nakhon Ratchasima 30000, Thailand;6. Photonics Technology Laboratory, National Electronics and Computer Technology Center (NECTEC), Pathumthani 12120, Thailand |
| Abstract: | 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. |
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