3种方法纯化兔抗H9N2亚型禽流感病毒血清的比较

Comparison of Purification of Rabbit Anti-H9N2 Subtype Avian Influenza Virus Serum by Three Methods

本试验分别用蛋白G亲和层析法、饱和硫酸铵法和辛酸-硫酸铵法纯化兔抗H9N2亚型禽流感病毒血清,发现蛋白G亲和层析法纯化得到的抗体纯度最高,回收率较低,抗体活性未变,操作简便,但成本较高,适合实验室研究及小量样品的纯化;饱和硫酸铵法纯化得到的抗体纯度较蛋白G亲和层析法低一些,抗体活性不变,可通过多次盐析来提高抗体纯度,多作为蛋白G亲和层析法纯化前的初步纯化;辛酸-硫酸铵法纯化得到的抗体纯度不及前两种方法,回收率较高,抗体活性亦无改变,纯化时对pH的精确性和辛酸的浓度要求较高,多用于样品的粗提和单抗的纯化。

Rabbit anti-H9N2 subtype avian influenza virus（AIV） serum was purified by protein G affinity chromatograph,caprylic acid-ammonium sulfate precipitation and saturated ammonium sulfate precipitation separately.It was found that the antibodies obtained from the first method was of the highest purity,unchanged activity,relatively low recovery rate,easy operation and high cost,which indicated that the first method was suitable for laboratory studies and purification in small sample volume.Antibodies obtained with the second method were of lower purity than that from the first method and of unchanged activity.Repeated salting out could improve the purity.It was suitable for preliminary purification before protein G affinity chromatograph.The third method was often used in crude extraction in large volume samples or the purification of ascitic fluid.With this method,purity of the antibodies obtained was lower than that from the other two methods,recovery rate was high and activity of antibodies unchanged.The purification demanded for relatively high accuracy of pH value and caprylic acid concentration.