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Competitive ELISA for the Measurement of Glycoprotein Purified from Acanthopanx senticosus  

Ha, Eun-Suk (Department of Food Science and Biotechnology, Kyonggi University)
Hwang, Soo-Hyun (Department of Food Science and Biotechnology, Kyonggi University)
Shin, Kwang-Soon (Department of Food Science and Biotechnology, Kyonggi University)
Yu, Kwang-Won (Department of Kimchi and Food Seience, National College of Science and Technology)
Lee, Keyung-Ho (Kolon Central Research Park)
Choi, Joo-Sun (Division of Metabolic Disease, Department of Biomedical Science, National Institute of Health)
Park, Woo-Mun (Research Team, GOOFOO Inc)
Yoon, Taek-Joon (Department of Food Science and Biotechnology, Kyonggi University)
Publication Information
Korean Journal of Food Science and Technology / v.35, no.6, 2003 , pp. 1209-1215 More about this Journal
Abstract
This study was carried out to establish a quantitative analysis method of separating immuno-activating substance (EN-SP) from Acanthopanax senticosus (A. senticosus) by competitive direct ELISA. Mouse antiserum (anti-EN-SP) against EN-SP was generated by immunization (s.c.) of EN-SP purified from A. senticosus as an immunogen. The titer of anti-EN-SP was about 1 : 400, and the optimal dilution of EN-SP-HRP conjugate was 1 : 1,000. When the standard curve was constructed by ELISA, its sensitivity was about $0.2{\mu}g/mL$. The coefficient variation of intra- and inter-assay were $6.13{\sim}8.81%$ and $6.73{\sim}8.60%$, respectively. According to the standard curve, the concentration of EN-SP in various senticosus extracts was found to be only $59.85\;{\mu}g$ in 10mg of extract from the bark of A. senticosus. Similarly, the immunostimulating activity to produce $TNF-{\alpha}$ or IL-12 among the various extracts of Acanthopanax was shown to be correlated with the content of EN-SP. These results demonstrated that competitive ELISA was a convenient, fast, reproducible, and accurate method for the determination of EN-SP as an immunologically active standard substance in extract of A. senticosus.
Keywords
Acanthopanax senticosus; competitive ELISA; cytokine;
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