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

Elsheikh, HM; Hamdy, GM; Ebeid, N. H.; Sallam, Kh. M.; El-Bayoumy, A. S. A.; Mehany, N. L.; Fathy, SA

doi:10.1080/15321819.2021.1993896

Cited by 1 publication

(2 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…HCG; was purified in our labs from pooled urine of pregnant women at the 10th week of gestation with high purity of 98.2% and biological activity (10,811 IU/mg) using alcohol precipitation, anion exchange, and gel chromatography according to our recently published method [11]. Polyclonal βHCG antisera; were produced from rabbits with high titer (1/104) and displacement (84.8%) using purified βHCG subunits according to our recently published method [11].…”

Section: Preparation Of Hcgmentioning

confidence: 99%

“…Specificity, the ability of the antibody to recognize an antigen even when the antigen is measured in a medium of similar substances that may be expected to be present [22]. The specificity was tested according to the method of Abraham [25], also called the 50% inhibition method, where specificity was calculated by dividing the concentration of HCG by the concentration of cross-reactant at % B/B0 equal to 50% to HCG standards, and the obtained specificity was 98.8% [11], indicating that the locally prepared polyclonal anti-βHCG specifically binds to HCG has no cross-reactivity with related glycoproteins.…”

Section: Hcg-ria Validationmentioning

confidence: 99%

See 1 more Smart Citation

Development of radioimmunoassay system for determination of human chorionic gonadotropin in human sera

Elsheikh

Fathy

Ebeid

et al. 2023

J Radioanal Nucl Chem

View full text Add to dashboard Cite

The main objective of this study was the preparation and evaluation of the primary reagents for the liquid phase human chorionic gonadotropin-radioimmunoassay (HCG-RIA) kit. Polyclonal antibody is specific and valid; monoclonal antibodies are more specific. In HCG-RIA technique, polyclonal antibodies for βHCG subunits with high binding and displacement % were used. 125I-HCG radioactive tracer was prepared with high yield 71.58 ± 0.92%, purity 99.2 ± 0.05%, and specific activity 170.42 ± 1.65 µCi/µg using chloramine T method, βHCG polyclonal antisera and HCG standards in assay buffer matrix were locally prepared. In-house HCG-RIA was developed with high sensitivity 0.5 mIU/ml, specificity (99%), precision (CV% < 6.4 and < 8.3 for intra-and inter-assay, respectively), and accuracy (recovery range 96.4–104.3%) could be used for quantitative estimation of HCG in human sera for monitoring pregnancy and diagnosis of gestational trophoblastic diseases.

show abstract

Section: Preparation Of Hcgmentioning

confidence: 99%