离心超滤法测定蛋白质标记物的放射化学纯度

张云艳; 万建美; 钱晓敏; 郭萍; 朱然; 张友九

doi:10.7538/hhx.2016.38.YX.2016061

离心超滤法测定蛋白质标记物的放射化学纯度

Determination on Radiochemical Purity of Labelled Protein With Centrifugal Ultrafiltration

摘要

摘要: 用高效液相色谱法(HPLC)、三氯醋酸(TCA)沉淀法和超滤法分离测定¹²⁵I-AAFP（抗过敏融合蛋白）标记物的放射化学纯度(RCP)，并对其进行比较。结果表明：三种方法均能有效分离标记物和游离碘，用超滤法、TCA沉淀法和HPLC三种方法测定¹²⁵I-AAFP低、中、高放射化学纯度，样本的RCP分别为0.62%±0.21%、7.25%±1.38%、0.97%±0.12%，61.58%±0.52%、63.18%±1.98%、63.77%±1.23%，98.89%±0.31%、98.76%±0.41%、98.80%±0.82%。当RCP很低时，超滤法与HPLC法测得的RCP间无明显差异(P>0.05)，而要低于TCA沉淀法(P<0.05)；除此之外，三种方法无明显差异(P>0.05)。这说明离心超滤法可用于分离测定蛋白质标记物的放射化学纯度。

Abstract: To explore the determination of radiochemical purity(RCP) of labelled protein with centifugal ultrafiltration, the radiochemical purity of ¹²⁵I-AAFP(anti-allergy fusion protein) was determined and compared by high performance liquid chromatography(HPLC), trichloroacetic acid(TCA) precipitation and centrifugal ultrafiltrations. It shows that three methods can separate the labelled ¹²⁵I-AAFP and free iodine-125 efficiently. The lower, middle and higher radiochemical purity of ¹²⁵I-AAFP which are determined by centrifugal ultrafiltration, TCA and HPLC respectively are 0.62%±0.21%, 7.25%±1.38% and 0.97%±0.12%; 61.58%±0.52%, 63.18%±1.98% and 63.77%±1.23%; 98.89%±0.31%, 98.76%±0.41% and 98.80%±0.82%. When RCP of ¹²⁵I-AAFP is very lower, the RCP measured with HPLC is not different from that with centrifugal ultrafiltration(P>0.05), whereas lower than that with TCA(P<0.05). Except this, there are no significant differences in the RCP of ¹²⁵I-AAFP determined with three different methods (P>0.05). Centrifugal ultrafiltrations can be used to determinate the radiochemical purity of labelled protein.

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