Purification and identification of anticancer proteins from Bidens alba against human colorectal carcinoma SW480 cell

• Main Authors: Huang, Chin-Wen, 黃靖雯
• Other Authors: Ong, Ping-Lin
• Format: Others
• Language: zh-TW
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/09344030847383428278

碩士 === 國立嘉義大學 === 生化科技學系研究所 === 99 === Recently, many studies of the anti-inflammatory, antioxidant activity, antihyperglycemic activity and anti-tumor activity of Bidens alba on human. The antiproliferative activity of the B. alba protein extracts were studied on human colorectal adencarcinoma SW480 cell line. The B. alba protein were extracted with 30 mM sodium hydrosulfite additive. The lightness of protein extract with enzymatic browning is increased by 20 fold, without losing of anti-cancer activity. The protein extract was precipitated with 35~65% saturated (NH4)2SO4, and re-dissolved in 10 mM PBS (pH 7.3) solution containing 7.5% PVPP. The lightness of solution increase 76 fold. The protein-extracted go through DEAE SepharoseTM ion exchanger chromatography (50 mM Tris-HCl, pH 7.5), and eluted with 210 mM NaCl. The purified proteins still have 95% anti-cancer activity. One group of proteins, with were 66~70 kDa molecular weight and had 63% anti-cancer activity, were got from DEAE SepharoseTM ion exchanger chromatography (50 mM Phosphate buffer, pH 6.5), SP SepharoseTM ion exchanger chromatography (50 mM Citric, pH 4) and gel slicing with native polyacrylamide gel electrophoresis. These results reveals that B. alba contain the protein with antiproliferation ability to SW480. These homogeneous toxin proteins help the future studies of the biochemical characterization and the protein-protein interaction with the membrane receptors of cancer cells.