Fishman Jordan B, Berg Eric A
Cold Spring Harb Protoc. 2018 Jun 1;2018(6):2018/6/pdb.prot099150. doi: 10.1101/pdb.prot099150.
Because IgY does not interact with either Protein A or Protein G, more traditional methods must be used for its isolation. To isolate chicken IgY from the yolk of chicken eggs, the yolks and the whites must be separated much in the way a cook does. Minimizing the egg white proteins is an important part of this process. Although eggs can be accumulated for short periods of time, IgY must be isolated from the yolk within 2-3 wk to avoid fouling of the material. In the first section of this protocol, sodium sulfate is used to precipitate protein fractions differentially from the chicken egg yolk. The second section describes an alternative differential precipitation technique that uses Polyethylene Glycol 6000 instead of sodium sulfate.
由于鸡免疫球蛋白(IgY)与蛋白A或蛋白G均不相互作用,因此必须采用更传统的方法来分离它。要从鸡蛋蛋黄中分离鸡IgY,必须像厨师那样将蛋黄和蛋清分开。尽量减少蛋清蛋白是这个过程的重要部分。虽然鸡蛋可以短时间储存,但必须在2至3周内从蛋黄中分离出IgY,以避免材料污染。在本方案的第一部分,使用硫酸钠从鸡蛋蛋黄中差异性沉淀蛋白质组分。第二部分描述了一种替代的差异性沉淀技术,该技术使用聚乙二醇6000代替硫酸钠。