Tini M, Jewell U R, Camenisch G, Chilov D, Gassmann M
Institute of Physiology, Winterthurerstr. 190, CH-8057 Zürich, Switzerland.
Comp Biochem Physiol A Mol Integr Physiol. 2002 Mar;131(3):569-74. doi: 10.1016/s1095-6433(01)00508-6.
Despite the fact that the use of chicken as immunization host brings many advantages to the production of polyclonal antibodies, the generation of egg yolk immunoglobulins (IgY) is rarely chosen. In this review, we report on the fast and efficient method for generation and affinity purification of IgY, in this case raised against the alpha-subunit of hypoxia-inducible factor-1 (HIF-1). The IgY antibody was successfully applied in a variety of methods and a number of different species for HIF-1alpha detection. In electrophoretic mobility shift assays, the IgY antibody recognized the native HIF-1 complex. The IgY antibody also detected HIF-1alpha protein on Western blots with extracts derived from human, monkey, pig, dog and mouse cell lines grown under hypoxic conditions. Immunofluorescence and immunoprecipitation experiments using the IgY antibody allowed detection and subcellular localization of HIF-1alpha in the nuclei of hypoxic cells. Chicken antibody production brings great benefit concerning the welfare of the immunized animals, due to non-invasive antibody harvesting with the added convenience of simple egg collection. An additional advantage is the fast and simple IgY isolation from egg yolk. IgY technology is a great improvement and should be considered as a good alternative to conventional polyclonal antibody production in mammals.
尽管将鸡用作免疫宿主在多克隆抗体生产方面具有诸多优势，但很少有人选择生产蛋黄免疫球蛋白（IgY）。在本综述中，我们报告了一种快速高效的IgY生成及亲和纯化方法，此次针对的是缺氧诱导因子-1（HIF-1）的α亚基。IgY抗体成功应用于多种方法及多个不同物种，用于检测HIF-1α。在电泳迁移率变动分析中，IgY抗体识别天然的HIF-1复合物。IgY抗体还能在蛋白质印迹法中检测来自在缺氧条件下培养的人、猴、猪、狗和小鼠细胞系提取物中的HIF-1α蛋白。使用IgY抗体进行的免疫荧光和免疫沉淀实验能够检测缺氧细胞细胞核中的HIF-1α并确定其亚细胞定位。鸡抗体生产在免疫动物福利方面带来了极大益处，因为抗体采集是非侵入性的，且收集鸡蛋操作简便。另一个优势是能从蛋黄中快速简便地分离IgY。IgY技术是一项重大改进，应被视为哺乳动物传统多克隆抗体制备的良好替代方法。
