Electricwala A
Division of Biotechnology, PHLS Centre of Applied Microbiology & Research, Porton Down, Salisbury, UK.
Bioseparation. 1994 Aug;4(4):263-9.
Human prolactin was partially purified from pituitary glands by a combination of alkaline extraction, ammonium sulphate precipitation, hydrophobic interaction chromatography and gel filtration. The early steps in the purification were carried out under stringent safety conditions to minimise the risk of exposure to potential viral pathogens present in human tissue. Alkaline extraction was carried out over a short time interval to limit possible deamidation of prolactin without loss in recovery. During chromatography on phenyl-Sepharose, it was found that bound prolactin could be eluted with aqueous buffer, in contrast to the organic solvents used by previous investigators. Subsequent gel filtration removed high molecular weight contaminant proteins and further confirmed the monomeric form of purified prolactin. This material could be further purified to near homogeneity by ion-exchange chromatography and other separation techniques. Some guidelines on the safety precautions undertaken during the extraction of prolactin are described.
通过碱提取、硫酸铵沉淀、疏水相互作用色谱和凝胶过滤相结合的方法，从垂体中部分纯化了人催乳素。纯化的早期步骤在严格的安全条件下进行，以将暴露于人体组织中潜在病毒病原体的风险降至最低。碱提取在短时间间隔内进行，以限制催乳素可能的脱酰胺化，同时不损失回收率。在苯基琼脂糖凝胶色谱过程中，发现与先前研究者使用的有机溶剂不同，结合的催乳素可用水性缓冲液洗脱。随后的凝胶过滤去除了高分子量的污染蛋白，并进一步证实了纯化催乳素的单体形式。该物质可通过离子交换色谱和其他分离技术进一步纯化至接近均一。文中描述了在催乳素提取过程中采取的一些安全预防措施指南。
