人绒毛膜促性腺激素的有效纯化和高特异性多克隆抗β hcg的生产作为放射免疫测定试剂盒的组成部分。

Q2 Health Professions

Journal of immunoassay & immunochemistry Pub Date : 2022-05-04 Epub Date: 2021-10-21 DOI:10.1080/15321819.2021.1993896

H M Elsheikh, G M Hamdy, N H Ebeid, Kh M Sallam, Asa El-Bayoumy, N L Mehany, S A Fathy

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引用次数: 1

摘要

本研究旨在通过DEAE Sepharose FF对人绒毛膜促性腺激素(HCG)进行简单捕获，并用Sephacryl S200 HR抛光，从孕妇尿液中纯化出生物活性高(10811 IU/mg)、纯度高(98.2%)的人绒毛膜促性腺激素(HCG)。所得的HCG通过SDS-PAGE进行了表征，并使用尿素处理方法解离成α和β亚基。将βHCG亚基注入家兔体内，制备高特异性的抗βHCG多克隆血清。该多克隆抗β hcg在家兔体内局部产生，并评估其结合滴度(1/10000)、位移(84.8%)和特异性(98.8%)。纯化的HCG与本地制备的多克隆抗βHCG抗血清作为内部放射免疫测定系统的基本组成部分，用于定量估计人血清中HCG的含量。

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Effective purification of human chorionic gonadotropin and production of highly specific polyclonal anti-βHCG as a component of radioimmunoassay kit.

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.

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来源期刊

Journal of immunoassay & immunochemistry 医学-免疫学

CiteScore

3.50

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： The Journal of Immunoassay & Immunochemistry is an international forum for rapid dissemination of research results and methodologies dealing with all aspects of immunoassay and immunochemistry, as well as selected aspects of immunology. They include receptor assay, enzyme-linked immunosorbent assay (ELISA) in all of its embodiments, ligand-based assays, biological markers of ligand-receptor interaction, in vivo and in vitro diagnostic reagents and techniques, diagnosis of AIDS, point-of-care testing, clinical immunology, antibody isolation and purification, and others.