le Darcel C
Can J Comp Med. 1978 Jul;42(3):370-2.
The recovery of viral antigen in a pure form from tissue can be hindered by the small amount of antigen present relative to the remaining normal tissue constituents with which the antigen may form loose associations. An antigen from the livers of chickens with erythroblastosis had earlier seemed to be associated with nucleic acid. The present work suggests that this material, which reacts with reagents for DNA and RNA but is soluble in 0.2N HClO4, is only a contaminant and not an integral part required for complete activity of the antigen. Treatment with 0.2N HClO4, together with precipitation of the active protein by 60% saturation of aqueous solutions with ammonium sulphate, removes the nucleic acid without destroying antigenicity. The procedure results in a marked degree of purification of the antigen (732-fold with respect to original liver protein) but contaminants still remain. A contaminant absorbing at 260 nm resisting extraction with 0.2N HClO4, can be partly eliminated if the solution is treated with ether-alcohol to remove lipid.
从组织中以纯净形式回收病毒抗原可能会受到阻碍，因为相对于其余正常组织成分而言，存在的抗原量很少，抗原可能会与这些成分形成松散的结合。早期，来自患成红细胞增多症鸡肝脏的一种抗原似乎与核酸有关。目前的研究表明，这种与DNA和RNA试剂发生反应但可溶于0.2N高氯酸的物质只是一种污染物，并非抗原完全活性所必需的组成部分。用0.2N高氯酸处理，再加上用硫酸铵使水溶液饱和度达到60%沉淀活性蛋白，可去除核酸而不破坏抗原性。该方法使抗原得到了显著程度的纯化（相对于原始肝脏蛋白而言提高了732倍），但仍有污染物残留。如果用乙醚 - 酒精处理溶液以去除脂质，在260nm处有吸收且能抵抗0.2N高氯酸提取的污染物可部分被去除。
