鹅卵黄抗体IgY的提纯及其酶标抗体的制备

Extraction and purification of IgY from goose egg yolk and preparation of rabbit anti-goose IgG labeled by HRP

本试验采用辛酸去脂、饱和硫酸铵溶液盐析法提纯鹅卵黄抗体Ig Y,采用SDS-PAGE方法分析其纯度;用纯化后的Ig Y免疫试验兔,收集血清,测定血清抗体效价;采用辛酸-硫酸铵法结合Protein G树脂亲和层析法纯化兔血清Ig G,用改良的高碘酸钠法进行辣根过氧化物酶标记,直接ELISA法测定标记物效价。结果表明,提纯的鹅卵黄Ig Y浓度为11.5 mg/m L,纯度约为92%;血清琼脂扩散抗体效价为1:32;Western Blotting试验证明,兔血清中含有抗鹅卵黄抗体Ig Y重链及轻链的特异性抗体;直接ELISA法测得标记物效价为1:1 100。

This experiment adopted the method of octanoic acid extraction, saturated ammonium sulfate solution to salting out and purify goose egg yolk antibody Ig Y and analyzed its purity by sodium sulphate polyacrylamide gel electrophoresis. Rabbits were inoculated with the purified Ig Y and seria were collected, the serum antibody titer was measured. The caprylic acid-ammonium combined with Protein G resin affinity chromatography were used to purify Ig G. The purified Ig G was labeled with HRP and the titer of labeled antibody was measured by direct ELISA method. The results showed that the purified goose egg yolk Ig Y reached the concentration of 11.5 mg/m L and purity about 92%.The titer of the antiserum was 1：32 by agar gel immunodiffusion test. Western blotting test indicated that the rabbit serum contained anti-goose egg yolk Ig Y with heavy chain and light chain of specific antibody. The purified Ig G was labeled with HRP and the titer of the antibody measured by direct ELISA method was 1：1 100.