有效纯化人绒毛膜促性腺激素，并制备高特异性的多克隆抗-βHCG，作为放射免疫试剂盒的一个组成部分。
Effective purification of human chorionic gonadotropin and production of highly specific polyclonal anti-βHCG as a component of radioimmunoassay kit.
机构信息
Department of Labeled Compounds, Hot Labs Centre, Atomic Energy Authority, Egypt.
Department of Biochemistry, Faculty of Science, Ain Shams University, Cairo, Egypt.
出版信息
J Immunoassay Immunochem. 2022 May 4;43(3):233-249. doi: 10.1080/15321819.2021.1993896. Epub 2021 Oct 21.
This study aimed to purify human chorionic gonadotropin (HCG) from the urine of pregnant women with high biological activity (10811 IU/mg) and purity (98.2%), by simple capturing of HCG using DEAE Sepharose FF and polishing using Sephacryl S200 HR. The HCG obtained was characterized by SDS-PAGE and dissociated into alpha and beta subunits using the urea treatment method. The βHCG subunits were injected into rabbits for the production of highly specific polyclonal anti-βHCG antisera. The polyclonal anti-βHCG was locally produced in rabbits and assessed for binding titer (1/10000), displacement (84.8%), and specificity (98.8%). Purified HCG along with locally prepared polyclonal anti-βHCG antisera were used as basic components of the in-house Radioimmunoassay system for quantitative estimation of HCG in human serum.
本研究旨在从生物活性高（10811IU/mg）、纯度高（98.2%）的孕妇尿液中提取人绒毛膜促性腺激素（HCG）。使用 DEAE Sepharose FF 简单捕获 HCG，并使用 Sephacryl S200 HR 进行抛光，从而获得 HCG。通过 SDS-PAGE 对获得的 HCG 进行特征分析，并使用脲处理方法将其解离为 α 和 β 亚基。将βHCG 亚基注入兔子体内，产生高度特异性的多克隆抗-βHCG 抗血清。在兔子体内局部产生多克隆抗-βHCG，并评估其结合效价（1/10000）、置换率（84.8%）和特异性（98.8%）。纯化的 HCG 与本地制备的多克隆抗-βHCG 抗血清一起，作为内部放射免疫测定系统的基本成分，用于定量估计人血清中的 HCG。
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