Ishikawa E, Yoshitake S
J Immunol Methods. 1980;38(1-2):117-23. doi: 10.1016/0022-1759(80)90336-1.
A mild procedure for the purification of rabbit Fab' antibodies is described. A small column of normal goat IgG-Sepharose 4B (3 cm x 5mm) was saturated with F(ab')2 prepared from rabbit anti-goat IgG, and anti-goat IgG Fab was eluted from the column by splitting the disulfide bond in the hinge of the F(ab')2 molecule using 10 or 12 mM 2-mercaptoethylamine at pH 7. The recovery of anti-goat IgG Fab' in the eluate was about 35% of anti-goat IgG F(ab')2 bound to the column, and the purity of anti-goat IgG Fab' eluted was more than 90%. This procedure may be applicable to most kinds of rabbit IgG antibodies and useful for various immunoassays.
本文描述了一种纯化兔Fab'抗体的温和方法。用一小柱正常山羊IgG-琼脂糖4B（3厘米×5毫米），用兔抗山羊IgG制备的F(ab')2使其饱和，然后通过在pH 7下用10或12 mM 2-巯基乙胺断裂F(ab')2分子铰链中的二硫键，从柱上洗脱抗山羊IgG Fab。洗脱液中抗山羊IgG Fab'的回收率约为结合到柱上的抗山羊IgG F(ab')2的35%，洗脱的抗山羊IgG Fab'的纯度超过90%。该方法可能适用于大多数种类的兔IgG抗体，对各种免疫测定有用。
