H M Elsheikh, G M Hamdy, N H Ebeid, Kh M Sallam, Asa El-Bayoumy, N L Mehany, S A Fathy
{"title":"Effective purification of human chorionic gonadotropin and production of highly specific polyclonal anti-βHCG as a component of radioimmunoassay kit.","authors":"H M Elsheikh, G M Hamdy, N H Ebeid, Kh M Sallam, Asa El-Bayoumy, N L Mehany, S A Fathy","doi":"10.1080/15321819.2021.1993896","DOIUrl":null,"url":null,"abstract":"<p><p>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.</p>","PeriodicalId":15990,"journal":{"name":"Journal of immunoassay & immunochemistry","volume":"43 3","pages":"233-249"},"PeriodicalIF":0.0000,"publicationDate":"2022-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of immunoassay & immunochemistry","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1080/15321819.2021.1993896","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2021/10/21 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"Health Professions","Score":null,"Total":0}
引用次数: 1
Abstract
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.
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.