Determination of antibodies to human growth hormone in serum by protein G affinity—reversed-phase tandem column chromatography with fluorescence detection

1991 ◽  
Vol 249 (1) ◽  
pp. 185-193 ◽  
Author(s):  
Alice Riggin ◽  
Fred E Regnier ◽  
J.Richard Sportsman
2017 ◽  
Vol 100 (4) ◽  
pp. 1023-1028
Author(s):  
Mamdouh R Rezk ◽  
Marwa F Mohamed ◽  
Faten Abdelaziz Fathalla ◽  
Mostafa A Shehata

Abstract A rapid and sensitive HPLC method was developed and validated for selective determination of the human growth hormone (hGH) somatropin in the presence of its deamidated and oxidized degradation products. Reversed-phase chromatography with an acetonitrile and ammonium bicarbonate mobile phase in gradient elution mode was used. A short run time of 15 min allowed rapid and cost-effective analysis, with an average retention time of 7.4 min for native hGH, 6.2 min for its deamidated form, and 4.3 and 6 min for its oxidized variants. The method was validated for selectivity, linearity, and intra- and interday variations according to International Conference on Harmonization guidelines. The proposed method was successfully applied for rapid evaluation of the quantity of hGH during downstream processing, formulation, and storage.


2004 ◽  
Vol 2004 (3) ◽  
pp. 143-149 ◽  
Author(s):  
Juliana F. Moura ◽  
Luiz DeLacerda ◽  
Romolo Sandrini ◽  
Fernanda M. Borba ◽  
Denise N. Castelo ◽  
...  

Human growth hormone (hGH) signal transduction initiates with a receptor dimerization in which one molecule binds to the receptor through sites 1 and 2. A sandwich enzyme-linked immunosorbent assay was developed for quantifying hGH molecules that present helix 4 from binding site 1. For this, horse anti-rhGH antibodies were eluted by an immunoaffinity column constituted by sepharose-rhGH. These antibodies were purified through a second column with synthetic peptide correspondent tohGH helix 4, immobilized to sepharose, and used as capture antibodies. Those that did not recognize synthetic peptide were used as a marker antibody. The working range was of 1.95 to 31.25 ng/mL of hGH. The intra-assay coefficient of variation (CV) was between 4.53% and 6.33%, while the interassay CV was between 6.00% and 8.27%. The recovery range was between 96.0% to 103.8%. There was no cross-reactivity with human prolactin. These features show that our assay is an efficient method for the determination of hGH.


The Analyst ◽  
2014 ◽  
Vol 139 (18) ◽  
pp. 4556-4563 ◽  
Author(s):  
V. Serafín ◽  
G. Martínez-García ◽  
L. Agüí ◽  
P. Yáñez-Sedeño ◽  
J. M. Pingarrón

A label-free dual electrochemical immunosensor for simultaneous determination of human growth and prolactin hormones was prepared for the first time.


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