疏水相互作用色谱法分离大豆凝集素

采用疏水相互作用色谱法（ＨＩＣ），以苯基琼脂糖凝胶ＳｅｐｈａｒｏｓｅＣＬ－４Ｂ为固定相，硫酸铵－磷酸盐缓冲溶液（ｐＨ６．８）为流动相，从大豆匀浆液中分离出大豆凝集素（ＳＢＡ）。

Separation of soybean agglutinin by hydrophobic-interaction chromatography]

The soybean agglutinin (SBA) was separated with hydrophobic-interaction chromatography (HIC) in this work. HIC of the crude lectin extract of SBA was performed on Phenyl-Sepharose CL-4B column, 10 cm x 2.5 cm i.d., at a flow-rate of 1.0 mL/min. A 20 mL sample of the crude extract was applied to the column previously equilibrated with the buffer, 0.01 mol/L PBS containing ammonium sulfate (60% of saturation concentration). The column was eluted with the starting buffer, then with the buffer containing 30% saturation of ammonium sulfate, and finally with 0.01 mol/L PBS. Fractions were detected by UV absorbance at 280 nm and assayed for hemagglutinating activity. The results show that the purification-fold was as high as 35 and the recovery of activity was 100%.