Huisman T H, Altay C, Webber B, Reese A L, Gravely M E, Okonjo K, Wilson J B
Blood. 1981 Jan;57(1):75-82.
A modification of a high pressure liquid chromatographic (HPLC) procedure is described that enables the complete separation and quantitation of the A gamma T, A gamma I, and G gamma chains in human fetal hemoglobin. The method, which is fast and accurate, requires 5 to 2000 micrograms Hb F. The purity of the Hb F is not essential and admixture of up to 70% adult Hb does not interfere with the determination. The method has been applied to the Hb F of 64 Black SS patients and 7 persons with the Hb S-HPFH (G gamma A gamma type) conditions. (A) Both "adult" G gamma to A gamma (2:3) and "newborn" G gamma to A gamma (3:2) ratios were observed in adult SS patients, 8 yr and older. Only 12% of the SS patients had the "newborn" ratio. This high G gamma to A gamma ratio may be due to a modification of the genetic switch mechanism that regulates the change of this ratio after birth. (B) Intermediate G gamma to A gamma ratios were only found in young SS patients, 5 yr of age or less. The results suggest a delayed switch of the newborn leads to adult ratio in sickle cell anemia. (C) The A gamma T chain was present in only 6% of all SS patients. One patient is homozygous for this variant chain. (D) Three of the 7 subjects with Hb S-HPFH were positive for the A gamma T chain. Its percentage was low, which suggests that the A gamma T chain gene is in trans of the HPFH determinant. (E) Quantitation of the three gamma chain types is also possible in the Hb F from Hb S heterozygotes with (nearly) normal Hb F levels. Such an analysis is useful for an evaluation of genetic conditions involving variations in the production of (different types of) Hb F.
本文描述了一种高压液相色谱（HPLC）方法的改进，该方法能够完全分离和定量人胎儿血红蛋白中的AγT、AγI和Gγ链。该方法快速且准确，所需血红蛋白F（Hb F）量为5至2000微克。Hb F的纯度并非必需，高达70%的成人血红蛋白混合也不影响测定。该方法已应用于64例黑人镰状细胞贫血（SS）患者和7例Hb S - HPFH（GγAγ型）患者的Hb F检测。（A）在8岁及以上的成人SS患者中，观察到了“成人型”Gγ与Aγ（2:3）以及“新生儿型”Gγ与Aγ（3:2）的比例。只有12%的SS患者具有“新生儿型”比例。这种高Gγ与Aγ比例可能是由于调节出生后该比例变化的基因开关机制发生了改变。（B）仅在5岁及以下的年轻SS患者中发现了中间的Gγ与Aγ比例。结果表明，新生儿向成人比例的转换延迟在镰状细胞贫血中出现。（C）AγT链仅在所有SS患者的6%中存在。有一名患者为该变异链的纯合子。（D）7例Hb S - HPFH患者中有3例AγT链呈阳性。其比例较低，这表明AγT链基因与HPFH决定簇处于反式状态。（E）对于（几乎）具有正常Hb F水平的Hb S杂合子的Hb F，也可以对三种γ链类型进行定量。这样的分析对于评估涉及（不同类型）Hb F产生变化的遗传状况很有用。
