Earticle

초록

영어

The purpose of this study is to optimize ELISA protocol to quantify colorectal cancer antigen GA733 and Fc antibody complex protein (GA733-Fc) expressed in transgenic plant. To confirm whether GA733 was functionally fused to Fc, mAb CO17-1A recognizing GA733 was coated on the 96 well ELISA plate. Goat anti-human IgG conjugated to horseradish peroxidase was applied to detect the Fc fragment of human IgG. ELISA was conducted by variable conditions of coating, washing, blocking, and sample amount. In coating, the mammalian-derived mAb CO17-1A were diluted at 2.5, 5, 10 μg/ml (100 μl/well). In washing, 1X PBS as compared to 1X PBS-T (1X PBS+0.05% Tween 20). In blocking, two types of buffers, 5% skim milk and 3% BSA solutions were compared each others. The results showed that 3% BSA gived better blocking than 5% skim milk. For plant samples, we applied transgenic plant leaf extracts expressing antigen GA733-Fc with dilution factors 1/2, 1/5, 1/10 and 1/100. Taken together, the optimized ELISA conditions to quantify plant-drived GA733-Fc were 0.5 μg of mAb CO17-1A per well for coating 3% BSA for blocking buffer, and 1/5 leaf extract sample dilution factor.

저자

• Junsik Ahn [ Department of Medicine, Medical Research Institute, College of Medicine, Chung-Ang University, Seoul 156-756 Republic of Korea ]
• Youngtae Byun [ Department of Medicine, Medical Research Institute, College of Medicine, Chung-Ang University, Seoul 156-756 Republic of Korea ]
• Jung-Hwan Lee [ Department of Medicine, Medical Research Institute, College of Medicine, Chung-Ang University, Seoul 156-756 Republic of Korea ]
• Deuk-Su Kim [ Department of Medicine, Medical Research Institute, College of Medicine, Chung-Ang University, Seoul 156-756 Republic of Korea ]
• Young-Kug Choo [ Department of Biological Science, College of Natural Sciences, Wonkwang University, Iksan, Jeonbuk 570-749, Republic of Korea ]
• Kyung-A Hwang [ Department of Agrofood Resources, National Academy of agricultural Science, Rural Development Administration, Suwon, Korea ]
• Sungsu Park [ Department of Chemistry and Nano Science Ewha Womans University, Seoul, Korea ]
• Kisung Ko [ Department of Medicine, Medical Research Institute, College of Medicine, Chung-Ang University, Seoul 156-756 Republic of Korea ]

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발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  한국당과학회 학술대회

• 간기

  연간

• 수록기간

  2006~2022

• 십진분류

  KDC 517 DDC 614