奥尼罗非鱼血清免疫球蛋白的纯化及分子量的初步分析

Purification and Determination of Molecular Weight in Serum Immunoglobulin from Tilapia

通过饱和硫酸铵盐析结合Sephadex G-200柱层析的方法,纯化制备了健康非免疫状态下奥尼罗非鱼的免疫球蛋白,进行变性还原条件下的聚丙烯酰胺凝胶电泳(SDS-PAGE)和免疫印迹试验对血清的免疫球蛋白进行初步分析,发现SDS-PAGE电泳条件下血清重链分子量为85 kD,轻链分子量为30 kD。如果罗非鱼血清免疫球蛋白在自然状态与其他硬骨鱼类一样也为四聚体,那么其总分子量的理论值应为920 kD。

Serum immunoglobulin（Ig） of healthy tilapia Oreochromis niloticus was purified by saturation ammonium sulfate precipitation and followed by Sephadex G -200 gel column chromatography. Molecular weight and partial characterization of heavy chain and light chain of Ig were measured by SDS polyacrylamide gel electrophoresis （ SDS - PAGE） and Western blotting. The results showed that the heavy chain had a molecular weight of 85 kD, and the light chain 30 kD. If the serum immunoglobulin（Ig） of tilapia is tetramer in nature ,the immunoglobulin would have a molecular weight of 920 kD.