摘要
A novel and efficient method for purification of soybean agglutinin (SBA) from soybean was reported. The method was characterized by selective extraction of SBA from soybean homogenate with barbiturate buffer (pH 6 2), removal of impurity by hydroxyapatite, and the final purification of SBA by guaran affinity chromatography. The purified SBA showed a single band of 27 5 kD by SDS PAGE. The lowest concentration of SBA that caused agglutination of the rabbit red blood cells was 0 31 mg/L. Agglutination of different cancer cell lines by the purified SBA was examined. Strong agglutination of the human nasopharyngeal CNE cells, mouse Lewis lung carcinoma cells, and rat mammary adenocarcinoma R3230AC cells was observed. However, SBA could not agglutinate the human hepatocellular carcinoma BEL 7402 cells, suggesting that unlike the above mentioned three cell lines, the BEL 7402 cells may not express N acetylgalactosamine (GalNAc) or galactose (Gal) residues in significant amount at the non reducing terminals of their cell surface glycans.
A novel and efficient method for purification of soybean agglutinin (SBA) from soybean was reported. The method was characterized by selective extraction of SBA from soybean homogenate with barbiturate buffer (pH 6 2), removal of impurity by hydroxyapatite, and the final purification of SBA by guaran affinity chromatography. The purified SBA showed a single band of 27 5 kD by SDS PAGE. The lowest concentration of SBA that caused agglutination of the rabbit red blood cells was 0 31 mg/L. Agglutination of different cancer cell lines by the purified SBA was examined. Strong agglutination of the human nasopharyngeal CNE cells, mouse Lewis lung carcinoma cells, and rat mammary adenocarcinoma R3230AC cells was observed. However, SBA could not agglutinate the human hepatocellular carcinoma BEL 7402 cells, suggesting that unlike the above mentioned three cell lines, the BEL 7402 cells may not express N acetylgalactosamine (GalNAc) or galactose (Gal) residues in significant amount at the non reducing terminals of their cell surface glycans.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2003年第3期401-405,共5页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金资助项目 (No .3 0 0 70 184)~~
关键词
大豆凝集素
纯化
肿瘤细胞
凝集反应
soybean agglutinin, affinity chromatography, hydroxyapatite, malignant cells