RAPID PROCEDURE FOR GLOBIN CHAIN ANALYSIS IN BLOOD-SAMPLES OF NORMAL AND BETA-THALASSEMIC FETUSES

  • 1 January 1981
    • journal article
    • research article
    • Vol. 57  (2) , 353-360
Abstract
Reversed-phase high pressure liquid chromatography (HPLC) was used for the separation of the .alpha., .beta., A.gamma. and G.gamma. and chains from human blood samples. The .alpha. and .beta. chains were normally eluted close together, but their separation was improved by coupling 2 or 3 columns in series, or by increasing the temperature of the columns. This method was applied for the determination of .beta./.gamma. ratios in blood samples obtained at fetoscopy from normal pregnancies and fetuses at risk for .beta.-thalassemia. The values obtained by high pressure chromatography were similar but slightly lower than those found by carboxymethyl cellulose (CMC) chromatography. The average G.gamma./A.gamma. ratio of the chains labeled after a 2 h pulse with [3H]leucine was almost identical to the actual G.gamma./A.gamma. measured by absorbance at 280 nm, indicating a constant rate of synthesis and accumulation of both globin chains in the 1st trimester fetus.