36B-18 |
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A. Y. Cho1, J. Y. PARK2, Y. M. Hur3, H. W. Lee4, J. Chung1, and S. Oh2. (1) Dept. of Biochemistry and Molecular Biology, Seoul National Univ. College of Medicine, 28 Yeongun-dong, Chongro-gu, Seoul, 110-799, South Korea, (2) Food and Nutritional Sciences, Ewha Womans Univ., 11-1 Daehyun-dong, Seodaemun-gu, Seoul, 120-750, South Korea, (3) Dept. of Biochemistry and Molecular Biology, Seoul National Univ. College of Medicine, 28 Yeongun-dong, Chongro-gu, Seoul, 110-799, South Korea, (4) Food and Nutritional Sciences, Ewha Womans University, 11-1 Deahyun-dong, Seodaemun-gu, Seoul, 120-750, South Korea Amygdalin (laterile, or vitamin B17) is a cyanogenic glycoside compound which commonly found in the pits of many fruits and raw nuts and in other plants, such as lima beans, clover, and sorghum. In the intestine, the enzyme beta-glucosidase present in gastrointestinal tract bacteria or many food plants hydrolyze amygdalin into cyanide which is absorbed into the lymph and portal circulations. As several plants contain amygdalin in sufficient amount to induce crucial cyanide poisoning in human, detecting amygdalin in food is sometime a life-saving measure. We immunized New Zealand White rabbits with Amygdalin-KLH (keyholelimpet hemocyanin) conjugate and succeeded to raise anti-sera reactive to amygdalin-ovalbumin conjugate. The sera did not show any reactivity to ovalbumin used for conjugation and the free amygdalin interfered the reactivity of sera to amydalin-ovalbumin conjugates. These data proved that amygdalin can behave as a hapten in rabbits. From bone marrow and spleen cDNA of immunized rabbit, phage display of combinatorial Fab and scFv library were constructed, of which complexity was over 1 X 109. We performed to select amygdalin-specific antibody clones from the library using various bio-panning procedures on amydalin-ovalbumin.
Session 36B, Biotechnology: General
2005 IFT Annual Meeting, July 15-20 - New Orleans, Louisiana |