抗HLA-A~*0201重链卵黄抗体的制备及初步鉴定

Preparation and identification of egg yolk antibodies against HLA-A~* 0201 heavy chain

目的制备高效价特异性鸡卵黄免疫球蛋白(Yolk immunoglobulin,IgY)抗体。方法用纯化HLA-A*0201重链抗原加完全福氏佐剂免疫岭南黄鸡,经水稀释法初步纯化浓缩后,用ELISA鉴定其免疫学活性,测定纯化后蛋白含量,并用SDS-PAGE进行分析鉴定。结果获得ELISA效价为1×106的抗重链IgY抗体,蛋白含量为9.77 mg/ml。经SDS-PAGE分析,出现2条蛋白带,纯度达90%以上。结论用重链抗原加福氏佐剂免疫种鸡制备的IgY抗体产量多、效价高,为结合HLA-A*0201复合体诱导特异性细胞毒性T淋巴细胞活化奠定了基础。

Objective To prepare a high-titer specific and efficient egg yolk immunoglobulin (IgY). Methods The antigen of HLA-A* 0201 heavy chain was used to immunize the hens together with Freud's adjuvant and the eggs these hens laid were collected. IgY was purified from the egg yolk by water extraction, salt precipitation with ammonium sulfate and dialysis. The immunoactivity of the IgY was measured by enzyme-linked immunosorbent assay (ELISA) and its purity was determined by SDS-PAGE. Results The titer of IgY was 1∶106 as shown by ELISA, with protein concentration of 9.77 mg/ml. The purity of the resultant IgY was more than 90% as suggested by SDS-PAGE analysis. Conclusion The antigen of HLA-A* 0201 heavy chain along with complete Freund adjuvant is effective to elicit an immune response of hens for producing IgY antibodies in high yields.