Randhawa Z I, Jones R T, Lie-Injo L E
Anal Biochem. 1983 Feb 15;129(1):184-91. doi: 10.1016/0003-2697(83)90067-2.
The zeta (zeta) and various gamma (gamma) chains of Hb Portland I have been separated and isolated from blood samples obtained from neonates with hydrops fetalis due to homozygous alpha-thalassemia. By using developers containing acetonitrile, methanol, and potassium phosphate and either an analytical (3.9 mm x 30 cm) or a preparative (7.8 mm x 30 cm) muBondapak C-18 column (Waters), globin chains from 200 micrograms to 5.0 mg have been isolated in pure forms. Analytical and preparative procedures using short (50-min duration) and extended (186-min duration) gradient programs have been developed. In addition to the type of column and developer conditions, the following factors are found to be important: (a) preparation of sample, (b) sample loading, and (c) cleaning of the column. Preliminary studies indicate that the yield ranges from 40 to 60% depending on the type of globin sample and the age of the column. This procedure also permits the separation of alpha, beta, and various gamma globin chains from fetal and adult samples.
已从因纯合子α地中海贫血导致胎儿水肿的新生儿血液样本中分离并提纯出波特兰血红蛋白I的ζ链和多种γ链。通过使用含有乙腈、甲醇和磷酸钾的展开剂，以及一根分析型（3.9毫米×30厘米）或制备型（7.8毫米×30厘米）的μBondapak C - 18柱（沃特世公司），已将200微克至5.0毫克的珠蛋白链提纯为纯净形式。现已开发出使用短梯度程序（50分钟时长）和长梯度程序（186分钟时长）的分析方法和制备方法。除了柱型和展开剂条件外，还发现以下因素很重要：（a）样品制备，（b）样品上样，以及（c）柱的清洗。初步研究表明，根据珠蛋白样品的类型和柱子的使用时长，产率在40%至60%之间。此方法还能从胎儿和成人样本中分离出α、β以及多种γ珠蛋白链。
