火箭免疫电泳标化蛋白质标准品中的白蛋白

CALIBRATION OF ALBUMIN IN PROTEIN STANDARD PREPARATION BY ELECTROIMMUNOASSAY

本文报道了用火箭免疫电泳标化蛋白质标准品中白蛋白的方法。白蛋白制品经95×1.4cm Sephadex G—200柱层析,取K_(AV)=0.45的白蛋白单体洗脱部分,用PAGE证实呈单一区带,浓缩后定值,以此作为标准进行白蛋白标化.探讨了该法的最适反应条件并进行了方法学的评价.对平均浓度为18.9、36.6、47.3g/L 的白蛋白样品进行重复性试验,批内变异系数分别为1.55%、1.45%和1.21%;批间变异为3.06%、2.5%和2.12%。回收率范围为97.6～101.8%。胆红素、血红蛋白、球蛋白及加入的防腐剂NaN_3不干扰标化.本文特异性强、准确性高、精密度好、有望成为蛋白质标准品中白蛋白标化的参考方法。

We describe a method for calibrating albumin in preparation of protein standard byelectroimmunoassay.Albumin preparation was chromatographed on a 95X1.4 cm column of Sephadex G-200.The fractions was pooled according to the expected K_(AV)value 0.45 for the monomer albumin.It gaved a singleband by polyacryamide gel electrophoresis.This purified monomer was used as calibration standard after concen-trating and determining the value of protein.We studied optimum condition of the reaction and evaluated themethodologic performance.With-run precision(CV)for the method was 1.55%,1.45% and 1.21% at meanalbumin concentration of 18.9,36.6 and 47.3 g/L,respectively;between-run CV was 3.06%,2.50%and 2.12%,respectively.Analytical recovery albumin added to pool(from 4.1 to 24.4 g/L)rangedfrom 97.6% to 101.6%.Bilirubin,hemoglobin,globulins and preservative sodium azide didn't interfere.Becauseof good precision,high accuracy and specificity,the method is a possible candidate reference method for cali-brating albumin.