抗鸡蛋黄IgY单克隆抗体的制备和初步鉴定

Preparation and Preliminary Characterization of Monoclonal Antibodies to Chicken Egg Yolk IgG

以纯化的鸡蛋黄ＩｇＧ（ＩｇＹ）为免疫原，免疫ＢＡＬＢ／ｃ小鼠。取其脾细胞与小鼠Ｓｐ２／０骨髓瘤细胞融合，经间接ＥＬＩＳＡ筛选，３次克隆化后，获得２株能稳定分泌抗ＩｇＹ单克隆抗体的杂交瘤细胞ＩＧ１１和３Ａ６。用琼脂双扩散法鉴定，两者均为ＩｇＧ１亚类。特异性鉴定表明，１Ｇ１１和３Ａ６。两株单克隆抗体均与鸡血清ＩｇＧ和鸡蛋黄ＩｇＧ起强反应，而不与鸭蛋黄、鹌鹑蛋黄、小鼠血清、大鼠血清、兔血清、羊血清、马血清，以及人血清的ＩｇＧ起反应。１Ｇ１１与３Ａ６的腹水效价分别为３．１×１０－６和６．４×１０－６。

Chicken egg yolk IgG(IgY) was used to immunize BALB/c mice and two hybridoma celllines, 1G11 and 3A6, were established by fusing Sp2/0 myeloma cells with the immunized mousespleen cells. By double gel diffusion test, 1G11 and 3A6 were shown to be IgG1. Both of theseMcAbs showed strong reaction with chicken sera IgG and egg yolk IgG, but no reaction with IgGsfrom duck egg yolk, quail egg yolk, mouse sera, rat sera, rabbit sera, goat sera, horse sera or humansera. The titers of McAb 1G11 and 3A6 from ascites were 3. 1 × 10- 6and 6. 4 × 10-6, respectively.