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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 high titer specific Yolk immunoglobulin (IgY) antibody. Methods Lingnan yellow chickens were immunized with purified HLA-A * 0201 heavy chain antigen plus complete Freund’s adjuvant. The purified chickens were purified by water dilution method and identified by ELISA. The purified protein was determined by SDS-PAGE Analysis and identification. Results An anti-heavy chain IgY antibody with ELISA titer of 1 × 106 was obtained with a protein content of 9.77 mg / ml. By SDS-PAGE analysis, there are two protein bands, the purity of more than 90%. Conclusion IgY antibody produced by immunized breeders with heavy chain antigen plus Freund ’s adjuvant has high yield and high titer, which lays the foundation for the induction of specific cytotoxic T lymphocyte activation by HLA-A * 0201 complex.