摘要
首次应用非连续聚丙烯酰胺凝胶(12％和7.5％)电泳将胚胎、胎儿及成人溶血液中各种正常珠蛋白链分离。其电泳移动顺序为:α、β、Gγ、δ、Aδ、ε和ζ链,δ与Gγ链得到较好的分离。10例β—地中海贫血杂合子血样的δ链光密扫描测定结果为2.62±0.76％。重型β—地中海贫血患者的Gγ与Aγ比1.09～1.27,各类样品的α/非α链含量比约为1。结合网织红细胞体外培育,~3H—亮氨酸标记新合成的珠蛋白链,以放射性荧光自显影技术测定了五例α—地贫患者的珠蛋白链合成速率,其α/β合成比均小于1(0.79～0.89),而正常对照为0.97。在珠蛋白链的研究中,该方法具有简单、灵敏等优点。
The discontinuous polyacrylamide gel(12% and 7.5%) electrophoresis (DPAGE) was designed and firstly applied to separate the human globin chains in embryonic, fetal and adult hemalysates. All known globin chains, including α, β, Gγ, σ, Aγ, ε and ξ, chains, were completely separated from one another. The a chain was the fastest band and the ξ chain was the slowest one among all globin chains in DPAGE. Especially, this method successed in separating the σ from the GY chains, which had not yet been reported. The amount of the 6 chain in hemalysates of 10 heterozygous β-thalassemia cases was determined by densitometry on DPAGE. It was 2.62±0.76% .Meanwhile, the level of HbA2in these cases measured by DE52 cellulose chromatography was 5.39±1.89%. It gave a good correlation coefficient of 0.97(P<0.001) between DPAGE and DE52 macrochromatography. The Gγ/Aγ ratios in hemalysates of homozygous β-thalassemia patients was 1.09-1.27. The results were different from the Gγ/Aγ ratios of newborn and adult. To diagnose thalassemia patients, reticulocytes were incubated with the medium contai ning 3H-Leucine. The labelled globin chains were separated by DPAGE and measured by autoradiofluorography. The α/β synthetic ratios in 5 α-thalassemia subjects were below 1 (0.79-0.89) and 0.97 in normal person. In general, this method is comparatively simple and sensitive for analysis of the human globin chains in he-malysates.
