Richardson P J, Siddle K, Luzio J P
Biochem J. 1984 Apr 15;219(2):647-54. doi: 10.1042/bj2190647.
A method for the immunoaffinity purification of cholinergic nerve terminals from mammalian brain was developed. A sheep antiserum to Torpedo electric-organ synaptic membranes, previously shown to be specific for cholinergic terminals in mammalian brain, was incubated with crude mitochondrial fractions prepared from rat brain. Cholinergic nerve terminals sensitized by this serum were purified from the mitochondrial fractions on a high-capacity cellulose immunoadsorbent bearing a mouse monoclonal anti-(sheep immunoglobulin G) antibody. Adsorption of nerve terminals on to the immunoadsorbent was assessed by using a variety of enzyme markers and gave a maximum yield of 24% of choline acetyltransferase, whereas non-specific binding was less than 1.0% for all of the enzymes measured. Cholinergic terminals were purified 26-fold from rat caudate nucleus, 30-fold from rat hippocampus and 38-fold from rat cerebral cortex. The terminals were shown to be intact, osmotically sensitive and metabolically active.
开发了一种从哺乳动物大脑中免疫亲和纯化胆碱能神经末梢的方法。将先前已证明对哺乳动物大脑中的胆碱能末梢具有特异性的抗电鳐电器官突触膜的绵羊抗血清与从大鼠大脑制备的粗线粒体组分一起孵育。通过带有小鼠单克隆抗（绵羊免疫球蛋白G）抗体的高容量纤维素免疫吸附剂，从线粒体组分中纯化被该血清致敏的胆碱能神经末梢。通过使用多种酶标记物评估神经末梢在免疫吸附剂上的吸附，胆碱乙酰转移酶的最大产率为24%，而对于所有测量的酶，非特异性结合均小于1.0%。胆碱能末梢从大鼠尾状核中纯化了26倍，从大鼠海马体中纯化了30倍，从大鼠大脑皮层中纯化了38倍。这些末梢显示是完整的、对渗透压敏感且具有代谢活性的。
