鸡卵黄免疫球蛋白的分离制备研究

Study on Seperation and Purification of Immunoglobulin in Yolk

实验研究了从高免鸡卵黄中分离纯化免疫球蛋白(IgY)的方法。确定最优条件为:卵黄十倍稀释,pH5.3,-18℃冻结6h,4℃解冻;添加3%硅藻土进行微滤;采用超滤浓缩五倍;再用45%饱和硫酸铵(pH5.3)盐析;沉淀重溶后经DEAE-Sepharose F.F.柱层析分离纯化。结果表明,用此方法可以从高免鸡蛋中得到电泳纯IgY,总活性回收率达到64.16%。

This experiment studied the method of separating and purifying immunoglobulin from the yolk of highly immuned hens, and established the optimized conditions: the yolk was diluted 10 times with aseptic water, regulated to pH 5.3, frozen at -18℃ for 6h, then thawed at 4℃. The thawed yolk solution was added with 3% diatomaceous earth, and filtered through 0. 45μm cellulose acetate membrane. Then the solution was concentrated 5 times through the way of Ultraflitration (5MWCO) and precipitated with 45% saturated ammonium sulfate under the condition of pH 5.3. In the end, it was separated and purified again through DEAE-Sepharose F.F. Column after the sediment was redissolved. The result showed that electrophoreticaly pure IgY could be obtained from the yolk of immuned hens and the Total Activity Recovery could reach 64.16%.